---
_id: '12082'
abstract:
- lang: eng
text: Proximity-dependent protein labeling provides a powerful in vivo strategy
to characterize the interactomes of specific proteins. We previously optimized
a proximity labeling protocol for Caenorhabditis elegans using the highly active
biotin ligase TurboID. A significant constraint on the sensitivity of TurboID
is the presence of abundant endogenously biotinylated proteins that take up bandwidth
in the mass spectrometer, notably carboxylases that use biotin as a cofactor.
In C. elegans, these comprise POD-2/acetyl-CoA carboxylase alpha, PCCA-1/propionyl-CoA
carboxylase alpha, PYC-1/pyruvate carboxylase, and MCCC-1/methylcrotonyl-CoA carboxylase
alpha. Here, we developed ways to remove these carboxylases prior to streptavidin
purification and mass spectrometry by engineering their corresponding genes to
add a C-terminal His10 tag. This allows us to deplete them from C. elegans lysates
using immobilized metal affinity chromatography. To demonstrate the method's efficacy,
we use it to expand the interactome map of the presynaptic active zone protein
ELKS-1. We identify many known active zone proteins, including UNC-10/RIM, SYD-2/liprin-alpha,
SAD-1/BRSK1, CLA-1/CLArinet, C16E9.2/Sentryn, as well as previously uncharacterized
potentially synaptic proteins such as the ortholog of human angiomotin, F59C12.3
and the uncharacterized protein R148.3. Our approach provides a quick and inexpensive
solution to a common contaminant problem in biotin-dependent proximity labeling.
The approach may be applicable to other model organisms and will enable deeper
and more complete analysis of interactors for proteins of interest.
acknowledged_ssus:
- _id: Bio
acknowledgement: "We thank de Bono laboratory members for helpful comments on the
article and the Mass Spec Facilities at IST Austria and Max Perutz Labs for invaluable
discussions and comments on how to optimize mass spec analyses of worm samples.
We are grateful to Ekaterina Lashmanova for designing the degron knock-in constructs
and preparing the injection mixes for CRISPR/Cas9-mediated genome editing. All LC–MS/MS
analyses were performed on instruments of the Vienna BioCenter Core Facilities instrument
pool.\r\nThis work was supported by a Wellcome Investigator Award (grant no.: 209504/Z/17/Z
) to M.d.B. and an ISTplus Fellowship to M.A. (Marie Sklodowska-Curie agreement
no.: 754411)."
article_number: '102343'
article_processing_charge: No
article_type: original
author:
- first_name: Murat
full_name: Artan, Murat
id: C407B586-6052-11E9-B3AE-7006E6697425
last_name: Artan
- first_name: Markus
full_name: Hartl, Markus
last_name: Hartl
- first_name: Weiqiang
full_name: Chen, Weiqiang
last_name: Chen
- first_name: Mario
full_name: De Bono, Mario
id: 4E3FF80E-F248-11E8-B48F-1D18A9856A87
last_name: De Bono
orcid: 0000-0001-8347-0443
citation:
ama: Artan M, Hartl M, Chen W, de Bono M. Depletion of endogenously biotinylated
carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in
Caenorhabditis elegans. Journal of Biological Chemistry. 2022;298(9). doi:10.1016/j.jbc.2022.102343
apa: Artan, M., Hartl, M., Chen, W., & de Bono, M. (2022). Depletion of endogenously
biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity
labeling in Caenorhabditis elegans. Journal of Biological Chemistry. Elsevier.
https://doi.org/10.1016/j.jbc.2022.102343
chicago: Artan, Murat, Markus Hartl, Weiqiang Chen, and Mario de Bono. “Depletion
of Endogenously Biotinylated Carboxylases Enhances the Sensitivity of TurboID-Mediated
Proximity Labeling in Caenorhabditis Elegans.” Journal of Biological Chemistry.
Elsevier, 2022. https://doi.org/10.1016/j.jbc.2022.102343.
ieee: M. Artan, M. Hartl, W. Chen, and M. de Bono, “Depletion of endogenously biotinylated
carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in
Caenorhabditis elegans,” Journal of Biological Chemistry, vol. 298, no.
9. Elsevier, 2022.
ista: Artan M, Hartl M, Chen W, de Bono M. 2022. Depletion of endogenously biotinylated
carboxylases enhances the sensitivity of TurboID-mediated proximity labeling in
Caenorhabditis elegans. Journal of Biological Chemistry. 298(9), 102343.
mla: Artan, Murat, et al. “Depletion of Endogenously Biotinylated Carboxylases Enhances
the Sensitivity of TurboID-Mediated Proximity Labeling in Caenorhabditis Elegans.”
Journal of Biological Chemistry, vol. 298, no. 9, 102343, Elsevier, 2022,
doi:10.1016/j.jbc.2022.102343.
short: M. Artan, M. Hartl, W. Chen, M. de Bono, Journal of Biological Chemistry
298 (2022).
date_created: 2022-09-11T22:01:55Z
date_published: 2022-09-01T00:00:00Z
date_updated: 2023-08-03T13:56:46Z
day: '01'
ddc:
- '570'
department:
- _id: MaDe
doi: 10.1016/j.jbc.2022.102343
ec_funded: 1
external_id:
isi:
- '000884241800011'
pmid:
- '35933017'
file:
- access_level: open_access
checksum: e726c7b9315230e6710e0b1f1d1677e9
content_type: application/pdf
creator: dernst
date_created: 2022-09-12T08:14:50Z
date_updated: 2022-09-12T08:14:50Z
file_id: '12092'
file_name: 2022_JBC_Artan.pdf
file_size: 2101656
relation: main_file
success: 1
file_date_updated: 2022-09-12T08:14:50Z
has_accepted_license: '1'
intvolume: ' 298'
isi: 1
issue: '9'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '09'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: 23870BE8-32DE-11EA-91FC-C7463DDC885E
grant_number: 209504/A/17/Z
name: Molecular mechanisms of neural circuit function
- _id: 260C2330-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '754411'
name: ISTplus - Postdoctoral Fellowships
publication: Journal of Biological Chemistry
publication_identifier:
eissn:
- 1083-351X
issn:
- 0021-9258
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Depletion of endogenously biotinylated carboxylases enhances the sensitivity
of TurboID-mediated proximity labeling in Caenorhabditis elegans
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 298
year: '2022'
...
---
_id: '10117'
abstract:
- lang: eng
text: Proximity labeling provides a powerful in vivo tool to characterize the proteome
of subcellular structures and the interactome of specific proteins. The nematode
Caenorhabditis elegans is one of the most intensely studied organisms in biology,
offering many advantages for biochemistry. Using the highly active biotin ligase
TurboID, we optimize here a proximity labeling protocol for C. elegans. An advantage
of TurboID is that biotin's high affinity for streptavidin means biotin-labeled
proteins can be affinity-purified under harsh denaturing conditions. By combining
extensive sonication with aggressive denaturation using SDS and urea, we achieved
near-complete solubilization of worm proteins. We then used this protocol to characterize
the proteomes of the worm gut, muscle, skin, and nervous system. Neurons are among
the smallest C. elegans cells. To probe the method's sensitivity, we expressed
TurboID exclusively in the two AFD neurons and showed that the protocol could
identify known and previously unknown proteins expressed selectively in AFD. The
active zones of synapses are composed of a protein matrix that is difficult to
solubilize and purify. To test if our protocol could solubilize active zone proteins,
we knocked TurboID into the endogenous elks-1 gene, which encodes a presynaptic
active zone protein. We identified many known ELKS-1-interacting active zone proteins,
as well as previously uncharacterized synaptic proteins. Versatile vectors and
the inherent advantages of using C. elegans, including fast growth and the ability
to rapidly make and functionally test knock-ins, make proximity labeling a valuable
addition to the armory of this model organism.
acknowledgement: We thank de Bono lab members for helpful comments on the manuscript,
IST Austria and University of Vienna Mass Spec Facilities for invaluable discussions
and comments for the optimization of mass spec analyses of worm samples. The biotin
auxotropic E. coli strain MG1655bioB:kan was gift from John Cronan (University of
Illinois) and was kindly sent to us by Jessica Feldman and Ariana Sanchez (Stanford
University). dg398 pEntryslot2_mNeongreen::3XFLAG::stop and dg397 pEntryslot3_mNeongreen::3XFLAG::stop::unc-54
3′UTR entry vector were kindly shared by Dr Dominique Glauser (University of Fribourg).
Codon-optimized mScarlet vector was a generous gift from Dr Manuel Zimmer (University
of Vienna).
article_number: '101094'
article_processing_charge: Yes
article_type: original
author:
- first_name: Murat
full_name: Artan, Murat
id: C407B586-6052-11E9-B3AE-7006E6697425
last_name: Artan
orcid: 0000-0001-8945-6992
- first_name: Stephen
full_name: Barratt, Stephen
id: 57740d2b-2a88-11ec-97cf-d9e6d1b39677
last_name: Barratt
- first_name: Sean M.
full_name: Flynn, Sean M.
last_name: Flynn
- first_name: Farida
full_name: Begum, Farida
last_name: Begum
- first_name: Mark
full_name: Skehel, Mark
last_name: Skehel
- first_name: Armel
full_name: Nicolas, Armel
id: 2A103192-F248-11E8-B48F-1D18A9856A87
last_name: Nicolas
- first_name: Mario
full_name: De Bono, Mario
id: 4E3FF80E-F248-11E8-B48F-1D18A9856A87
last_name: De Bono
orcid: 0000-0001-8347-0443
citation:
ama: Artan M, Barratt S, Flynn SM, et al. Interactome analysis of Caenorhabditis
elegans synapses by TurboID-based proximity labeling. Journal of Biological
Chemistry. 2021;297(3). doi:10.1016/J.JBC.2021.101094
apa: Artan, M., Barratt, S., Flynn, S. M., Begum, F., Skehel, M., Nicolas, A., &
de Bono, M. (2021). Interactome analysis of Caenorhabditis elegans synapses by
TurboID-based proximity labeling. Journal of Biological Chemistry. Elsevier.
https://doi.org/10.1016/J.JBC.2021.101094
chicago: Artan, Murat, Stephen Barratt, Sean M. Flynn, Farida Begum, Mark Skehel,
Armel Nicolas, and Mario de Bono. “Interactome Analysis of Caenorhabditis Elegans
Synapses by TurboID-Based Proximity Labeling.” Journal of Biological Chemistry.
Elsevier, 2021. https://doi.org/10.1016/J.JBC.2021.101094.
ieee: M. Artan et al., “Interactome analysis of Caenorhabditis elegans synapses
by TurboID-based proximity labeling,” Journal of Biological Chemistry,
vol. 297, no. 3. Elsevier, 2021.
ista: Artan M, Barratt S, Flynn SM, Begum F, Skehel M, Nicolas A, de Bono M. 2021.
Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity
labeling. Journal of Biological Chemistry. 297(3), 101094.
mla: Artan, Murat, et al. “Interactome Analysis of Caenorhabditis Elegans Synapses
by TurboID-Based Proximity Labeling.” Journal of Biological Chemistry,
vol. 297, no. 3, 101094, Elsevier, 2021, doi:10.1016/J.JBC.2021.101094.
short: M. Artan, S. Barratt, S.M. Flynn, F. Begum, M. Skehel, A. Nicolas, M. de
Bono, Journal of Biological Chemistry 297 (2021).
date_created: 2021-10-10T22:01:23Z
date_published: 2021-09-01T00:00:00Z
date_updated: 2023-08-14T07:24:09Z
day: '01'
ddc:
- '612'
department:
- _id: MaDe
- _id: LifeSc
doi: 10.1016/J.JBC.2021.101094
ec_funded: 1
external_id:
isi:
- '000706409200006'
file:
- access_level: open_access
checksum: 19e39d36c5b9387c6dc0e89c9ae856ab
content_type: application/pdf
creator: cchlebak
date_created: 2021-10-11T12:20:58Z
date_updated: 2021-10-11T12:20:58Z
file_id: '10121'
file_name: 2021_JBC_Artan.pdf
file_size: 1680010
relation: main_file
success: 1
file_date_updated: 2021-10-11T12:20:58Z
has_accepted_license: '1'
intvolume: ' 297'
isi: 1
issue: '3'
language:
- iso: eng
month: '09'
oa: 1
oa_version: Published Version
project:
- _id: 260C2330-B435-11E9-9278-68D0E5697425
call_identifier: H2020
grant_number: '754411'
name: ISTplus - Postdoctoral Fellowships
publication: Journal of Biological Chemistry
publication_identifier:
eissn:
- 1083-351X
issn:
- 0021-9258
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Interactome analysis of Caenorhabditis elegans synapses by TurboID-based proximity
labeling
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 297
year: '2021'
...
---
_id: '8440'
abstract:
- lang: eng
text: Mycobacterium tuberculosis can remain dormant in the host, an ability that
explains the failure of many current tuberculosis treatments. Recently, the natural
products cyclomarin, ecumicin, and lassomycin have been shown to efficiently kill
Mycobacterium tuberculosis persisters. Their target is the N-terminal domain of
the hexameric AAA+ ATPase ClpC1, which recognizes, unfolds, and translocates protein
substrates, such as proteins containing phosphorylated arginine residues, to the
ClpP1P2 protease for degradation. Surprisingly, these antibiotics do not inhibit
ClpC1 ATPase activity, and how they cause cell death is still unclear. Here, using
NMR and small-angle X-ray scattering, we demonstrate that arginine-phosphate binding
to the ClpC1 N-terminal domain induces millisecond dynamics. We show that these
dynamics are caused by conformational changes and do not result from unfolding
or oligomerization of this domain. Cyclomarin binding to this domain specifically
blocked these N-terminal dynamics. On the basis of these results, we propose a
mechanism of action involving cyclomarin-induced restriction of ClpC1 dynamics,
which modulates the chaperone enzymatic activity leading eventually to cell death.
article_processing_charge: No
article_type: original
author:
- first_name: Katharina
full_name: Weinhäupl, Katharina
last_name: Weinhäupl
- first_name: Martha
full_name: Brennich, Martha
last_name: Brennich
- first_name: Uli
full_name: Kazmaier, Uli
last_name: Kazmaier
- first_name: Joel
full_name: Lelievre, Joel
last_name: Lelievre
- first_name: Lluis
full_name: Ballell, Lluis
last_name: Ballell
- first_name: Alfred
full_name: Goldberg, Alfred
last_name: Goldberg
- first_name: Paul
full_name: Schanda, Paul
id: 7B541462-FAF6-11E9-A490-E8DFE5697425
last_name: Schanda
orcid: 0000-0002-9350-7606
- first_name: Hugo
full_name: Fraga, Hugo
last_name: Fraga
citation:
ama: Weinhäupl K, Brennich M, Kazmaier U, et al. The antibiotic cyclomarin blocks
arginine-phosphate–induced millisecond dynamics in the N-terminal domain of ClpC1
from Mycobacterium tuberculosis. Journal of Biological Chemistry. 2018;293(22):8379-8393.
doi:10.1074/jbc.ra118.002251
apa: Weinhäupl, K., Brennich, M., Kazmaier, U., Lelievre, J., Ballell, L., Goldberg,
A., … Fraga, H. (2018). The antibiotic cyclomarin blocks arginine-phosphate–induced
millisecond dynamics in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis.
Journal of Biological Chemistry. American Society for Biochemistry &
Molecular Biology. https://doi.org/10.1074/jbc.ra118.002251
chicago: Weinhäupl, Katharina, Martha Brennich, Uli Kazmaier, Joel Lelievre, Lluis
Ballell, Alfred Goldberg, Paul Schanda, and Hugo Fraga. “The Antibiotic Cyclomarin
Blocks Arginine-Phosphate–Induced Millisecond Dynamics in the N-Terminal Domain
of ClpC1 from Mycobacterium Tuberculosis.” Journal of Biological Chemistry.
American Society for Biochemistry & Molecular Biology, 2018. https://doi.org/10.1074/jbc.ra118.002251.
ieee: K. Weinhäupl et al., “The antibiotic cyclomarin blocks arginine-phosphate–induced
millisecond dynamics in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis,”
Journal of Biological Chemistry, vol. 293, no. 22. American Society for
Biochemistry & Molecular Biology, pp. 8379–8393, 2018.
ista: Weinhäupl K, Brennich M, Kazmaier U, Lelievre J, Ballell L, Goldberg A, Schanda
P, Fraga H. 2018. The antibiotic cyclomarin blocks arginine-phosphate–induced
millisecond dynamics in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis.
Journal of Biological Chemistry. 293(22), 8379–8393.
mla: Weinhäupl, Katharina, et al. “The Antibiotic Cyclomarin Blocks Arginine-Phosphate–Induced
Millisecond Dynamics in the N-Terminal Domain of ClpC1 from Mycobacterium Tuberculosis.”
Journal of Biological Chemistry, vol. 293, no. 22, American Society for
Biochemistry & Molecular Biology, 2018, pp. 8379–93, doi:10.1074/jbc.ra118.002251.
short: K. Weinhäupl, M. Brennich, U. Kazmaier, J. Lelievre, L. Ballell, A. Goldberg,
P. Schanda, H. Fraga, Journal of Biological Chemistry 293 (2018) 8379–8393.
date_created: 2020-09-18T10:05:18Z
date_published: 2018-06-01T00:00:00Z
date_updated: 2021-01-12T08:19:17Z
day: '01'
doi: 10.1074/jbc.ra118.002251
extern: '1'
intvolume: ' 293'
issue: '22'
keyword:
- Cell Biology
- Biochemistry
- Molecular Biology
language:
- iso: eng
month: '06'
oa_version: None
page: 8379-8393
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
- 1083-351X
publication_status: published
publisher: American Society for Biochemistry & Molecular Biology
quality_controlled: '1'
status: public
title: The antibiotic cyclomarin blocks arginine-phosphate–induced millisecond dynamics
in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 293
year: '2018'
...
---
_id: '6298'
abstract:
- lang: eng
text: Tumor necrosis factor-stimulated gene-6 (TSG-6) is a hyalu-ronan (HA)-binding
protein that plays important roles ininflammation and ovulation. TSG-6-mediated
cross-linking ofHA has been proposed as a functional mechanism (e.g.for regu-lating
leukocyte adhesion), but direct evidence for cross-linkingis lacking, and we know
very little about its impact on HA ultra-structure. Here we used films of polymeric
and oligomeric HAchains, end-grafted to a solid support, and a combination ofsurface-sensitive
biophysical techniques to quantify the bindingof TSG-6 into HA films and to correlate
binding to morpholog-ical changes. We find that full-length TSG-6 binds with pro-nounced
positive cooperativity and demonstrate that it cancross-link HA at physiologically
relevant concentrations. Ourdata indicate that cooperative binding of full-length
TSG-6arises from HA-induced protein oligomerization and that theTSG-6 oligomers
act as cross-linkers. In contrast, the HA-bind-ing domain of TSG-6 (the Link module)
alone binds withoutpositive cooperativity and weaker than the full-length protein.Both
the Link module and full-length TSG-6 condensed andrigidified HA films, and the
degree of condensation scaled withthe affinity between the TSG-6 constructs and
HA. We proposethat condensation is the result of protein-mediated HA cross-linking.
Our findings firmly establish that TSG-6 is a potent HAcross-linking agent and
might hence have important implica-tions for the mechanistic understanding of
the biological func-tion of TSG-6 (e.g.in inflammation).
author:
- first_name: Natalia
full_name: Baranova, Natalia
id: 38661662-F248-11E8-B48F-1D18A9856A87
last_name: Baranova
orcid: 0000-0002-3086-9124
- first_name: Erik
full_name: Nilebäck, Erik
last_name: Nilebäck
- first_name: F. Michael
full_name: Haller, F. Michael
last_name: Haller
- first_name: David C.
full_name: Briggs, David C.
last_name: Briggs
- first_name: Sofia
full_name: Svedhem, Sofia
last_name: Svedhem
- first_name: Anthony J.
full_name: Day, Anthony J.
last_name: Day
- first_name: Ralf P.
full_name: Richter, Ralf P.
last_name: Richter
citation:
ama: Baranova NS, Nilebäck E, Haller FM, et al. The inflammation-associated protein
TSG-6 cross-links hyaluronan via hyaluronan-induced TSG-6 oligomers. Journal
of Biological Chemistry. 2011;286(29):25675-25686. doi:10.1074/jbc.m111.247395
apa: Baranova, N. S., Nilebäck, E., Haller, F. M., Briggs, D. C., Svedhem, S., Day,
A. J., & Richter, R. P. (2011). The inflammation-associated protein TSG-6
cross-links hyaluronan via hyaluronan-induced TSG-6 oligomers. Journal of Biological
Chemistry. American Society for Biochemistry & Molecular Biology. https://doi.org/10.1074/jbc.m111.247395
chicago: Baranova, Natalia S., Erik Nilebäck, F. Michael Haller, David C. Briggs,
Sofia Svedhem, Anthony J. Day, and Ralf P. Richter. “The Inflammation-Associated
Protein TSG-6 Cross-Links Hyaluronan via Hyaluronan-Induced TSG-6 Oligomers.”
Journal of Biological Chemistry. American Society for Biochemistry &
Molecular Biology, 2011. https://doi.org/10.1074/jbc.m111.247395.
ieee: N. S. Baranova et al., “The inflammation-associated protein TSG-6 cross-links
hyaluronan via hyaluronan-induced TSG-6 oligomers,” Journal of Biological Chemistry,
vol. 286, no. 29. American Society for Biochemistry & Molecular Biology, pp.
25675–25686, 2011.
ista: Baranova NS, Nilebäck E, Haller FM, Briggs DC, Svedhem S, Day AJ, Richter
RP. 2011. The inflammation-associated protein TSG-6 cross-links hyaluronan via
hyaluronan-induced TSG-6 oligomers. Journal of Biological Chemistry. 286(29),
25675–25686.
mla: Baranova, Natalia S., et al. “The Inflammation-Associated Protein TSG-6 Cross-Links
Hyaluronan via Hyaluronan-Induced TSG-6 Oligomers.” Journal of Biological Chemistry,
vol. 286, no. 29, American Society for Biochemistry & Molecular Biology, 2011,
pp. 25675–86, doi:10.1074/jbc.m111.247395.
short: N.S. Baranova, E. Nilebäck, F.M. Haller, D.C. Briggs, S. Svedhem, A.J. Day,
R.P. Richter, Journal of Biological Chemistry 286 (2011) 25675–25686.
date_created: 2019-04-11T20:57:43Z
date_published: 2011-07-22T00:00:00Z
date_updated: 2021-01-12T08:06:58Z
day: '22'
doi: 10.1074/jbc.m111.247395
extern: '1'
intvolume: ' 286'
issue: '29'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: http://www.jbc.org/content/286/29/25675.full.pdf
month: '07'
oa: 1
oa_version: Published Version
page: 25675-25686
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
- 1083-351X
publication_status: published
publisher: American Society for Biochemistry & Molecular Biology
quality_controlled: '1'
status: public
title: The inflammation-associated protein TSG-6 cross-links hyaluronan via hyaluronan-induced
TSG-6 oligomers
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 286
year: '2011'
...
---
_id: '8473'
abstract:
- lang: eng
text: β2-microglobulin (β2m), the light chain of class I major histocompatibility
complex, is responsible for the dialysis-related amyloidosis and, in patients
undergoing long term dialysis, the full-length and chemically unmodified β2m converts
into amyloid fibrils. The protein, belonging to the immunoglobulin superfamily,
in common to other members of this family, experiences during its folding a long-lived
intermediate associated to the trans-to-cis isomerization of Pro-32 that has been
addressed as the precursor of the amyloid fibril formation. In this respect, previous
studies on the W60G β2m mutant, showing that the lack of Trp-60 prevents fibril
formation in mild aggregating condition, prompted us to reinvestigate the refolding
kinetics of wild type and W60G β2m at atomic resolution by real-time NMR. The
analysis, conducted at ambient temperature by the band selective flip angle short
transient real-time two-dimensional NMR techniques and probing the β2m states
every 15 s, revealed a more complex folding energy landscape than previously reported
for wild type β2m, involving more than a single intermediate species, and shedding
new light into the fibrillogenic pathway. Moreover, a significant difference in
the kinetic scheme previously characterized by optical spectroscopic methods was
discovered for the W60G β2m mutant.
article_processing_charge: No
article_type: original
author:
- first_name: Alessandra
full_name: Corazza, Alessandra
last_name: Corazza
- first_name: Enrico
full_name: Rennella, Enrico
last_name: Rennella
- first_name: Paul
full_name: Schanda, Paul
id: 7B541462-FAF6-11E9-A490-E8DFE5697425
last_name: Schanda
orcid: 0000-0002-9350-7606
- first_name: Maria Chiara
full_name: Mimmi, Maria Chiara
last_name: Mimmi
- first_name: Thomas
full_name: Cutuil, Thomas
last_name: Cutuil
- first_name: Sara
full_name: Raimondi, Sara
last_name: Raimondi
- first_name: Sofia
full_name: Giorgetti, Sofia
last_name: Giorgetti
- first_name: Federico
full_name: Fogolari, Federico
last_name: Fogolari
- first_name: Paolo
full_name: Viglino, Paolo
last_name: Viglino
- first_name: Lucio
full_name: Frydman, Lucio
last_name: Frydman
- first_name: Maayan
full_name: Gal, Maayan
last_name: Gal
- first_name: Vittorio
full_name: Bellotti, Vittorio
last_name: Bellotti
- first_name: Bernhard
full_name: Brutscher, Bernhard
last_name: Brutscher
- first_name: Gennaro
full_name: Esposito, Gennaro
last_name: Esposito
citation:
ama: Corazza A, Rennella E, Schanda P, et al. Native-unlike long-lived intermediates
along the folding pathway of the amyloidogenic protein β2-Microglobulin revealed
by real-time two-dimensional NMR. Journal of Biological Chemistry. 2010;285(8):5827-5835.
doi:10.1074/jbc.m109.061168
apa: Corazza, A., Rennella, E., Schanda, P., Mimmi, M. C., Cutuil, T., Raimondi,
S., … Esposito, G. (2010). Native-unlike long-lived intermediates along the folding
pathway of the amyloidogenic protein β2-Microglobulin revealed by real-time two-dimensional
NMR. Journal of Biological Chemistry. American Society for Biochemistry
& Molecular Biology. https://doi.org/10.1074/jbc.m109.061168
chicago: Corazza, Alessandra, Enrico Rennella, Paul Schanda, Maria Chiara Mimmi,
Thomas Cutuil, Sara Raimondi, Sofia Giorgetti, et al. “Native-Unlike Long-Lived
Intermediates along the Folding Pathway of the Amyloidogenic Protein Β2-Microglobulin
Revealed by Real-Time Two-Dimensional NMR.” Journal of Biological Chemistry.
American Society for Biochemistry & Molecular Biology, 2010. https://doi.org/10.1074/jbc.m109.061168.
ieee: A. Corazza et al., “Native-unlike long-lived intermediates along the
folding pathway of the amyloidogenic protein β2-Microglobulin revealed by real-time
two-dimensional NMR,” Journal of Biological Chemistry, vol. 285, no. 8.
American Society for Biochemistry & Molecular Biology, pp. 5827–5835, 2010.
ista: Corazza A, Rennella E, Schanda P, Mimmi MC, Cutuil T, Raimondi S, Giorgetti
S, Fogolari F, Viglino P, Frydman L, Gal M, Bellotti V, Brutscher B, Esposito
G. 2010. Native-unlike long-lived intermediates along the folding pathway of the
amyloidogenic protein β2-Microglobulin revealed by real-time two-dimensional NMR.
Journal of Biological Chemistry. 285(8), 5827–5835.
mla: Corazza, Alessandra, et al. “Native-Unlike Long-Lived Intermediates along the
Folding Pathway of the Amyloidogenic Protein Β2-Microglobulin Revealed by Real-Time
Two-Dimensional NMR.” Journal of Biological Chemistry, vol. 285, no. 8,
American Society for Biochemistry & Molecular Biology, 2010, pp. 5827–35,
doi:10.1074/jbc.m109.061168.
short: A. Corazza, E. Rennella, P. Schanda, M.C. Mimmi, T. Cutuil, S. Raimondi,
S. Giorgetti, F. Fogolari, P. Viglino, L. Frydman, M. Gal, V. Bellotti, B. Brutscher,
G. Esposito, Journal of Biological Chemistry 285 (2010) 5827–5835.
date_created: 2020-09-18T10:11:23Z
date_published: 2010-02-19T00:00:00Z
date_updated: 2021-01-12T08:19:31Z
day: '19'
doi: 10.1074/jbc.m109.061168
extern: '1'
intvolume: ' 285'
issue: '8'
keyword:
- Cell Biology
- Biochemistry
- Molecular Biology
language:
- iso: eng
month: '02'
oa_version: None
page: 5827-5835
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
- 1083-351X
publication_status: published
publisher: American Society for Biochemistry & Molecular Biology
quality_controlled: '1'
status: public
title: Native-unlike long-lived intermediates along the folding pathway of the amyloidogenic
protein β2-Microglobulin revealed by real-time two-dimensional NMR
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 285
year: '2010'
...
---
_id: '2614'
abstract:
- lang: eng
text: Metabotropic glutamate receptors (mGluRs) from group III reduce glutamate
release. Because these receptors reduce cAMP levels, we explored whether this
signaling pathway contributes to release inhibition caused by mGluRs with low
affinity for L-2-amino-4-phosphonobutyrate (L-AP4). In biochemical experiments
with the population of cerebrocortical nerve terminals we find that L-AP4 (1 mM)
inhibited the Ca2+dependent-evoked release of glutamate by 25%. This inhibitory
effect was largely prevented by the pertussis toxin but was insensitive to inhibitors
of protein kinase C bisindolylmaleimide and protein kinase A H-89. Furthermore,
this inhibition was associated with reduction in N-type Ca2+ channel activity
in the absence of any detectable change in cAMP levels. In the presence of forskolin,
however, L-AP4 decreased the levels of cAMP. The activation of this additional
signaling pathway was very efficient in counteracting the facilitation of glutamate
release induced either by forskolin or the β-adrenergic receptor agonist isoproterenol.
Imaging experiments to measure Ca2+ dynamics in single nerve terminals showed
that L-AP4 strongly reduced the Ca2+ response in 28% of the nerve terminals. Moreover,
immunochemical experiments showed that 25-35% of the nerve terminals that were
immunopositive to synaptophysin were also immunoreactive to the low affinity L-AP4-sensitive
mGluR7. Then, mGluR7 mediates the inhibition of glutamate release caused by 1
mM L-AP4, primarily by a strong inhibition of Ca2+ channels, although high cAMP
uncovers the receptor ability to decrease cAMP.
acknowledgement: We thank Dr. Enrique Castro from Las Palmas University for critical
reading of the manuscript and M. Sefton for editorial assistance.
article_processing_charge: No
article_type: original
author:
- first_name: Carmelo
full_name: Millán, Carmelo
last_name: Millán
- first_name: Rafael
full_name: Luján, Rafael
last_name: Luján
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: José
full_name: Sánchez Prieto, José
last_name: Sánchez Prieto
citation:
ama: Millán C, Luján R, Shigemoto R, Sánchez Prieto J. The inhibition of glutamate
release by metabotropic glutamate receptor 7 affects both [Ca2+]c and cAMP. Evidence
for a strong reduction of Ca2+ entry in single nerve terminals. Journal of
Biological Chemistry. 2002;277(16):14092-14101. doi:10.1074/jbc.M109044200
apa: Millán, C., Luján, R., Shigemoto, R., & Sánchez Prieto, J. (2002). The
inhibition of glutamate release by metabotropic glutamate receptor 7 affects both
[Ca2+]c and cAMP. Evidence for a strong reduction of Ca2+ entry in single nerve
terminals. Journal of Biological Chemistry. American Society for Biochemistry
and Molecular Biology. https://doi.org/10.1074/jbc.M109044200
chicago: Millán, Carmelo, Rafael Luján, Ryuichi Shigemoto, and José Sánchez Prieto.
“The Inhibition of Glutamate Release by Metabotropic Glutamate Receptor 7 Affects
Both [Ca2+]c and CAMP. Evidence for a Strong Reduction of Ca2+ Entry in Single
Nerve Terminals.” Journal of Biological Chemistry. American Society for
Biochemistry and Molecular Biology, 2002. https://doi.org/10.1074/jbc.M109044200.
ieee: C. Millán, R. Luján, R. Shigemoto, and J. Sánchez Prieto, “The inhibition
of glutamate release by metabotropic glutamate receptor 7 affects both [Ca2+]c
and cAMP. Evidence for a strong reduction of Ca2+ entry in single nerve terminals,”
Journal of Biological Chemistry, vol. 277, no. 16. American Society for
Biochemistry and Molecular Biology, pp. 14092–14101, 2002.
ista: Millán C, Luján R, Shigemoto R, Sánchez Prieto J. 2002. The inhibition of
glutamate release by metabotropic glutamate receptor 7 affects both [Ca2+]c and
cAMP. Evidence for a strong reduction of Ca2+ entry in single nerve terminals.
Journal of Biological Chemistry. 277(16), 14092–14101.
mla: Millán, Carmelo, et al. “The Inhibition of Glutamate Release by Metabotropic
Glutamate Receptor 7 Affects Both [Ca2+]c and CAMP. Evidence for a Strong Reduction
of Ca2+ Entry in Single Nerve Terminals.” Journal of Biological Chemistry,
vol. 277, no. 16, American Society for Biochemistry and Molecular Biology, 2002,
pp. 14092–101, doi:10.1074/jbc.M109044200.
short: C. Millán, R. Luján, R. Shigemoto, J. Sánchez Prieto, Journal of Biological
Chemistry 277 (2002) 14092–14101.
date_created: 2018-12-11T11:58:41Z
date_published: 2002-04-19T00:00:00Z
date_updated: 2023-07-25T10:16:44Z
day: '19'
ddc:
- '570'
doi: 10.1074/jbc.M109044200
extern: '1'
external_id:
pmid:
- '11825890'
file:
- access_level: open_access
checksum: 0290fcbbd9153ec654185b0c856f214c
content_type: application/pdf
creator: alisjak
date_created: 2023-07-25T10:13:16Z
date_updated: 2023-07-25T10:13:16Z
file_id: '13309'
file_name: 2002_JBC_Millan.pdf
file_size: 2105520
relation: main_file
success: 1
file_date_updated: 2023-07-25T10:13:16Z
has_accepted_license: '1'
intvolume: ' 277'
issue: '16'
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
page: 14092 - 14101
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4284'
quality_controlled: '1'
scopus_import: '1'
status: public
title: The inhibition of glutamate release by metabotropic glutamate receptor 7 affects
both [Ca2+]c and cAMP. Evidence for a strong reduction of Ca2+ entry in single nerve
terminals
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 277
year: '2002'
...
---
_id: '2621'
abstract:
- lang: eng
text: The release properties of glutamatergic nerve terminals are influenced by
a number of factors, including the subtype of voltage-dependent calcium channel
and the presence of presynaptic autoreceptors. Group III metabotropic glutamate
receptors (mGluRs) mediate feedback inhibition of glutamate release by inhibiting
Ca2+ channel activity. By imaging Ca2+ in preparations of cerebrocortical nerve
terminals, we show that voltage-dependent Ca2+ channels are distributed in a heterogeneous
manner in individual nerve terminals. Presynaptic terminals contained only N-type
(47.5%; conotoxin GVIA-sensitive), P/Q-type (3.9%; agatoxin IVA-sensitive), or
both N- and P/Q-type (42.6%) Ca2+ channels, although the remainder of the terminals
(6.1%) were insensitive to these two toxins. In this preparation, two mGluRs with
high and low affinity for L(+)-2-amino-4-phosphonobutyrate were identified by
immunocytochemistry as mGluR4 and mGluR7, respectively. These receptors were responsible
for 22.2 and 24.1% reduction of glutamate release, and they reduced the Ca2+ response
in 24.4 and 30.3% of the nerve terminals, respectively. Interestingly, mGluR4
was largely (73.7%) located in nerve terminals expressing both N- and P/Q-type
Ca2+ channels, whereas mGluR7 was predominantly (69.9%) located in N-type Ca2+
channel-expressing terminals. This specific coexpression of different group III
mGluRs and Ca2+ channels may endow synaptic terminals with distinct release properties
and reveals the existence of a high degree of presynaptic heterogeneity.
acknowledgement: We thank M. Sefton for editorial assistance.
article_processing_charge: No
article_type: original
author:
- first_name: Carmelo
full_name: Millán, Carmelo
last_name: Millán
- first_name: Rafael
full_name: Luján, Rafael
last_name: Luján
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: José
full_name: Sánchez Prieto, José
last_name: Sánchez Prieto
citation:
ama: Millán C, Luján R, Shigemoto R, Sánchez Prieto J. Subtype-specific expression
of Group III metabotropic glutamate receptors and Ca2+ channels in single nerve
terminals. Journal of Biological Chemistry. 2002;277(49):47796-47803. doi:10.1074/jbc.M207531200
apa: Millán, C., Luján, R., Shigemoto, R., & Sánchez Prieto, J. (2002). Subtype-specific
expression of Group III metabotropic glutamate receptors and Ca2+ channels in
single nerve terminals. Journal of Biological Chemistry. American Society
for Biochemistry and Molecular Biology. https://doi.org/10.1074/jbc.M207531200
chicago: Millán, Carmelo, Rafael Luján, Ryuichi Shigemoto, and José Sánchez Prieto.
“Subtype-Specific Expression of Group III Metabotropic Glutamate Receptors and
Ca2+ Channels in Single Nerve Terminals.” Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology, 2002. https://doi.org/10.1074/jbc.M207531200.
ieee: C. Millán, R. Luján, R. Shigemoto, and J. Sánchez Prieto, “Subtype-specific
expression of Group III metabotropic glutamate receptors and Ca2+ channels in
single nerve terminals,” Journal of Biological Chemistry, vol. 277, no.
49. American Society for Biochemistry and Molecular Biology, pp. 47796–47803,
2002.
ista: Millán C, Luján R, Shigemoto R, Sánchez Prieto J. 2002. Subtype-specific expression
of Group III metabotropic glutamate receptors and Ca2+ channels in single nerve
terminals. Journal of Biological Chemistry. 277(49), 47796–47803.
mla: Millán, Carmelo, et al. “Subtype-Specific Expression of Group III Metabotropic
Glutamate Receptors and Ca2+ Channels in Single Nerve Terminals.” Journal of
Biological Chemistry, vol. 277, no. 49, American Society for Biochemistry
and Molecular Biology, 2002, pp. 47796–803, doi:10.1074/jbc.M207531200.
short: C. Millán, R. Luján, R. Shigemoto, J. Sánchez Prieto, Journal of Biological
Chemistry 277 (2002) 47796–47803.
date_created: 2018-12-11T11:58:43Z
date_published: 2002-12-02T00:00:00Z
date_updated: 2023-07-19T07:49:19Z
day: '02'
doi: 10.1074/jbc.M207531200
extern: '1'
external_id:
pmid:
- '12376542'
intvolume: ' 277'
issue: '49'
language:
- iso: eng
month: '12'
oa_version: Published Version
page: 47796 - 47803
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4277'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Subtype-specific expression of Group III metabotropic glutamate receptors and
Ca2+ channels in single nerve terminals
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 277
year: '2002'
...
---
_id: '13438'
abstract:
- lang: eng
text: ICln is an ion channel identified by expression cloning using a cDNA library
from Madin-Darby canine kidney cells. In all organisms tested so far, only one
transcript for the ICln protein could be identified. Here we show that two splice
variants of the ICln ion channel can be found in Caenorhabditis elegans. Moreover,
we show that these two splice variants of the ICln channel protein, which we termed
IClnN1 and IClnN2, can be functionally reconstituted and tested in an artificial
lipid bilayer. In these experiments, the IClnN1-induced currents showed no voltage-dependent
inactivation, whereas the IClnN2-induced currents fully inactivated at positive
potentials. The molecular entity responsible for the voltage-dependent inactivation
of IClnN2 is a cluster of positively charged amino acids encoded by exon 2a, which
is absent in IClnN1. Our experiments suggest a mechanism of channel inactivation
that is similar to the “ball and chain” model proposed for the Shaker potassium
channel,i.e. a cluster of positively charged amino acids hinders ion permeation
through the channel by a molecular and voltage-dependent interaction at the inner
vestibulum of the pore. This hypothesis is supported by the finding that synthetic
peptides with the same amino acid sequence as the positive cluster can transform
the IClnN1-induced current to the current observed after reconstitution of IClnN2.
Furthermore, we show that the nematode ICln gene is embedded in an operon harboring
two additional genes, which we termed Nx and Ny. Co-reconstitution of Nx and IClnN2
and functional analysis of the related currents revealed a functional interaction
between the two proteins, as evidenced by the fact that the IClnN2-induced current
in the presence of Nx was no longer voltage-sensitive. The experiments described
indicate that the genome organization in nematodes allows an effective approach
for the identification of functional partner proteins of ion channels.
acknowledgement: We are grateful to D. E. Clapham, E. Wöll, G. Meyer, and G. Botta
for helpful discussion and/or reading of the manuscript. We also thank T. Stiernagle
for providing the N2 strain of C. elegans and A. Wimmer and M. Frick for technical
assistance
article_processing_charge: No
article_type: original
author:
- first_name: Johannes
full_name: Fürst, Johannes
last_name: Fürst
- first_name: Markus
full_name: Ritter, Markus
last_name: Ritter
- first_name: Jakob
full_name: Rudzki, Jakob
last_name: Rudzki
- first_name: Johann G
full_name: Danzl, Johann G
id: 42EFD3B6-F248-11E8-B48F-1D18A9856A87
last_name: Danzl
orcid: 0000-0001-8559-3973
- first_name: Martin
full_name: Gschwentner, Martin
last_name: Gschwentner
- first_name: Elke
full_name: Scandella, Elke
last_name: Scandella
- first_name: Martin
full_name: Jakab, Martin
last_name: Jakab
- first_name: Matthias
full_name: König, Matthias
last_name: König
- first_name: Bernhard
full_name: Oehl, Bernhard
last_name: Oehl
- first_name: Florian
full_name: Lang, Florian
last_name: Lang
- first_name: Peter
full_name: Deetjen, Peter
last_name: Deetjen
- first_name: Markus
full_name: Paulmichl, Markus
last_name: Paulmichl
citation:
ama: Fürst J, Ritter M, Rudzki J, et al. ICln Ion channel splice variants in Caenorhabditis
elegans. Journal of Biological Chemistry. 2002;277(6):4435-4445. doi:10.1074/jbc.m107372200
apa: Fürst, J., Ritter, M., Rudzki, J., Danzl, J. G., Gschwentner, M., Scandella,
E., … Paulmichl, M. (2002). ICln Ion channel splice variants in Caenorhabditis
elegans. Journal of Biological Chemistry. Elsevier. https://doi.org/10.1074/jbc.m107372200
chicago: Fürst, Johannes, Markus Ritter, Jakob Rudzki, Johann G Danzl, Martin Gschwentner,
Elke Scandella, Martin Jakab, et al. “ICln Ion Channel Splice Variants in Caenorhabditis
Elegans.” Journal of Biological Chemistry. Elsevier, 2002. https://doi.org/10.1074/jbc.m107372200.
ieee: J. Fürst et al., “ICln Ion channel splice variants in Caenorhabditis
elegans,” Journal of Biological Chemistry, vol. 277, no. 6. Elsevier, pp.
4435–4445, 2002.
ista: Fürst J, Ritter M, Rudzki J, Danzl JG, Gschwentner M, Scandella E, Jakab M,
König M, Oehl B, Lang F, Deetjen P, Paulmichl M. 2002. ICln Ion channel splice
variants in Caenorhabditis elegans. Journal of Biological Chemistry. 277(6), 4435–4445.
mla: Fürst, Johannes, et al. “ICln Ion Channel Splice Variants in Caenorhabditis
Elegans.” Journal of Biological Chemistry, vol. 277, no. 6, Elsevier, 2002,
pp. 4435–45, doi:10.1074/jbc.m107372200.
short: J. Fürst, M. Ritter, J. Rudzki, J.G. Danzl, M. Gschwentner, E. Scandella,
M. Jakab, M. König, B. Oehl, F. Lang, P. Deetjen, M. Paulmichl, Journal of Biological
Chemistry 277 (2002) 4435–4445.
date_created: 2023-08-01T12:37:50Z
date_published: 2002-02-08T00:00:00Z
date_updated: 2023-08-01T12:55:54Z
day: '08'
ddc:
- '570'
doi: 10.1074/jbc.m107372200
extern: '1'
external_id:
pmid:
- '11706026'
file:
- access_level: open_access
checksum: 13abe20f78eb37ab62beb006f62c69b7
content_type: application/pdf
creator: alisjak
date_created: 2023-08-01T12:44:09Z
date_updated: 2023-08-01T12:44:09Z
file_id: '13439'
file_name: 2002_JBC_Fuerst.pdf
file_size: 798920
relation: main_file
success: 1
file_date_updated: 2023-08-01T12:44:09Z
has_accepted_license: '1'
intvolume: ' 277'
issue: '6'
keyword:
- Cell Biology
- Molecular Biology
- Biochemistry
language:
- iso: eng
month: '02'
oa: 1
oa_version: Published Version
page: 4435-4445
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: ICln Ion channel splice variants in Caenorhabditis elegans
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 277
year: '2002'
...
---
_id: '3928'
abstract:
- lang: eng
text: Regulated adhesion of leukocytes to the extracellular matrix is essential
for transmigration of blood vessels and subsequent migration into the stroma of
inflamed tissues. Although beta(2)-integrins play an indisputable role in adhesion
of polymorphonuclear granulocytes (PMN) to endothelium, we show here that beta(1)-
and beta(3)-integrins but not beta(2)-integrin are essential for the adhesion
to and migration on extracellular matrix molecules of the endothelial cell basement
membrane and subjacent interstitial matrix. Mouse wild type and beta(2)-integrin
null PMN and the progranulocytic cell line 32DC13 were employed in in vitro adhesion
and migration assays using extracellular matrix molecules expressed at sites of
extravasation in vivo, in particular the endothelial cell laminins 8 and 10. Wild
type and beta(2)-integrin null PMN showed the same pattern of ECM binding, indicating
that beta(2)-integrins do not mediate specific adhesion of PMN to the extracellular
matrix molecules tested; binding was observed to the interstitial matrix molecules,
fibronectin and vitronectin, via integrins alpha(5)beta(1) and alpha(v)beta(3),
respectively; to laminin 10 via alpha(6)beta(1); but not to laminins 1, 2, and
8, collagen type I and IV, perlecan, or tenascin-C. PMN binding to laminins 1,
2, and 8 could not be induced despite surface expression of functionally active
integrin alpha(6)beta(1), a major laminin receptor, demonstrating that expression
of alpha(6)beta(1) alone is insufficient for ligand binding and suggesting the
involvement of accessory factors. Nevertheless, laminins 1, 8, and 10 supported
PMN migration, indicating that differential cellular signaling via laminins is
independent of the extent of adhesion. The data demonstrate that adhesive and
nonadhesive interactions with components of the endothelial cell basement membrane
and subjacent interstitium play decisive roles in controlling PMN movement into
sites of inflammation and illustrate that beta(2)-integrins are not essential
for such interactions.
acknowledgement: We thank Dr. T. Winkler for carrying out flow cytometry analysis,
Dr. Simon Goodman for providing cyclic RGD peptides and helpful discussions, and
Stefanie Karosi and Thomas Samson for critical review of the manuscript. This work
would not have been possible without the expert technical assistance of Friederike
Pausch.
article_processing_charge: No
article_type: original
author:
- first_name: Michael K
full_name: Sixt, Michael K
id: 41E9FBEA-F248-11E8-B48F-1D18A9856A87
last_name: Sixt
orcid: 0000-0002-6620-9179
- first_name: Rupert
full_name: Hallmann, Rupert
last_name: Hallmann
- first_name: Olaf
full_name: Wendler, Olaf
last_name: Wendler
- first_name: Karin
full_name: Scharffetter Kochanek, Karin
last_name: Scharffetter Kochanek
- first_name: Lydia
full_name: Sorokin, Lydia
last_name: Sorokin
citation:
ama: Sixt MK, Hallmann R, Wendler O, Scharffetter Kochanek K, Sorokin L. Cell adhesion
and migration properties of β2-integrin negative polymorphonuclear granulocytes
on defined extracellular matrix molecules. Relevance for leukocyte extravasation.
Journal of Biological Chemistry. 2001;276(22):18878-18887. doi:10.1074/jbc.M010898200
apa: Sixt, M. K., Hallmann, R., Wendler, O., Scharffetter Kochanek, K., & Sorokin,
L. (2001). Cell adhesion and migration properties of β2-integrin negative polymorphonuclear
granulocytes on defined extracellular matrix molecules. Relevance for leukocyte
extravasation. Journal of Biological Chemistry. American Society for Biochemistry
and Molecular Biology. https://doi.org/10.1074/jbc.M010898200
chicago: Sixt, Michael K, Rupert Hallmann, Olaf Wendler, Karin Scharffetter Kochanek,
and Lydia Sorokin. “Cell Adhesion and Migration Properties of Β2-Integrin Negative
Polymorphonuclear Granulocytes on Defined Extracellular Matrix Molecules. Relevance
for Leukocyte Extravasation.” Journal of Biological Chemistry. American
Society for Biochemistry and Molecular Biology, 2001. https://doi.org/10.1074/jbc.M010898200.
ieee: M. K. Sixt, R. Hallmann, O. Wendler, K. Scharffetter Kochanek, and L. Sorokin,
“Cell adhesion and migration properties of β2-integrin negative polymorphonuclear
granulocytes on defined extracellular matrix molecules. Relevance for leukocyte
extravasation,” Journal of Biological Chemistry, vol. 276, no. 22. American
Society for Biochemistry and Molecular Biology, pp. 18878–18887, 2001.
ista: Sixt MK, Hallmann R, Wendler O, Scharffetter Kochanek K, Sorokin L. 2001.
Cell adhesion and migration properties of β2-integrin negative polymorphonuclear
granulocytes on defined extracellular matrix molecules. Relevance for leukocyte
extravasation. Journal of Biological Chemistry. 276(22), 18878–18887.
mla: Sixt, Michael K., et al. “Cell Adhesion and Migration Properties of Β2-Integrin
Negative Polymorphonuclear Granulocytes on Defined Extracellular Matrix Molecules.
Relevance for Leukocyte Extravasation.” Journal of Biological Chemistry,
vol. 276, no. 22, American Society for Biochemistry and Molecular Biology, 2001,
pp. 18878–87, doi:10.1074/jbc.M010898200.
short: M.K. Sixt, R. Hallmann, O. Wendler, K. Scharffetter Kochanek, L. Sorokin,
Journal of Biological Chemistry 276 (2001) 18878–18887.
date_created: 2018-12-11T12:05:56Z
date_published: 2001-06-01T00:00:00Z
date_updated: 2023-05-11T12:54:06Z
day: '01'
doi: 10.1074/jbc.M010898200
extern: '1'
external_id:
pmid:
- '11278780'
intvolume: ' 276'
issue: '22'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925819670134?via%3Dihub
month: '06'
oa: 1
oa_version: Published Version
page: 18878 - 18887
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '2199'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Cell adhesion and migration properties of β2-integrin negative polymorphonuclear
granulocytes on defined extracellular matrix molecules. Relevance for leukocyte
extravasation
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 276
year: '2001'
...
---
_id: '3149'
abstract:
- lang: eng
text: The prohormone convertases (PCs) are an evolutionarily ancient group of proteases
required for the maturation of neuropeptide and peptide hormone precursors. In
Drosophila melanogaster, the homolog of prohormone convertase 2, dPC2 (amontillado),
is required for normal hatching behavior, and immunoblotting data indicate that
flies express 80- and 75-kDa forms of this protein. Because mouse PC2 (mPC2) requires
7B2, a helper protein for productive maturation, we searched the fly data base
for the 7B2 signature motif PPNPCP and identified an expressed sequence tag clone
encoding the entire open reading frame for this protein. dPC2 and d7B2 cDNAs were
subcloned into expression vectors for transfection into HEK-293 cells; mPC2 and
rat 7B2 were used as controls. Although active mPC2 was detected in medium in
the presence of either d7B2 or r7B2, dPC2 showed no proteolytic activity upon
coexpression of either d7B2 or r7B2. Labeling experiments showed that dPC2 was
synthesized but not secreted from HEK-293 cells. However, when dPC2 and either
d7B2 or r7B2 were coexpressed in Drosophila S2 cells, abundant immunoreactive
dPC2 was secreted into the medium, coincident with the appearance of PC2 activity.
Expression and secretion of dPC2 enzyme activity thus appears to require insect
cell-specific posttranslational processing events. The significant differences
in the cell biology of the insect and mammalian enzymes, with 7B2 absolutely required
for secretion of dPC2 and zymogen conversion occurring intracellularly in the
case of dPC2 but not mPC2, support the idea that the Drosophila enzyme has specific
requirements for maturation and secretion that can be met only in insect cells.
acknowledgement: This work was supported by National Institutes of Health Grants DK49703
(to I. L.), NS21749 (to P. H. T.), and GM39697 (to R. S. F.). The costs of publication
of this article were defrayed in part by the payment of page charges. This article
must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section
1734 solely to indicate this fact. 10749852. We thank members of the Lindberg laboratory
and Laurent Muller for helpful comments, Bin Tu for construction of the C. elegans
PC2 expression vector, and Joelle Finley for assistance with cell culture.
article_processing_charge: No
article_type: original
author:
- first_name: Jae
full_name: Hwang, Jae
last_name: Hwang
- first_name: Daria E
full_name: Siekhaus, Daria E
id: 3D224B9E-F248-11E8-B48F-1D18A9856A87
last_name: Siekhaus
orcid: 0000-0001-8323-8353
- first_name: Robert
full_name: Fuller, Robert
last_name: Fuller
- first_name: Paul
full_name: Taghert, Paul
last_name: Taghert
- first_name: Iris
full_name: Lindberg, Iris
last_name: Lindberg
citation:
ama: 'Hwang J, Siekhaus DE, Fuller R, Taghert P, Lindberg I. Interaction of Drosophila
melanogaster prohormone convertase 2 and 7B2: Insect cell specific processing
and secretion. Journal of Biological Chemistry. 2000;275(23):17886-17893.
doi:10.1074/jbc.M000032200 '
apa: 'Hwang, J., Siekhaus, D. E., Fuller, R., Taghert, P., & Lindberg, I. (2000).
Interaction of Drosophila melanogaster prohormone convertase 2 and 7B2: Insect
cell specific processing and secretion. Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology. https://doi.org/10.1074/jbc.M000032200 '
chicago: 'Hwang, Jae, Daria E Siekhaus, Robert Fuller, Paul Taghert, and Iris Lindberg.
“Interaction of Drosophila Melanogaster Prohormone Convertase 2 and 7B2: Insect
Cell Specific Processing and Secretion.” Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology, 2000. https://doi.org/10.1074/jbc.M000032200 .'
ieee: 'J. Hwang, D. E. Siekhaus, R. Fuller, P. Taghert, and I. Lindberg, “Interaction
of Drosophila melanogaster prohormone convertase 2 and 7B2: Insect cell specific
processing and secretion,” Journal of Biological Chemistry, vol. 275, no.
23. American Society for Biochemistry and Molecular Biology, pp. 17886–17893,
2000.'
ista: 'Hwang J, Siekhaus DE, Fuller R, Taghert P, Lindberg I. 2000. Interaction
of Drosophila melanogaster prohormone convertase 2 and 7B2: Insect cell specific
processing and secretion. Journal of Biological Chemistry. 275(23), 17886–17893.'
mla: 'Hwang, Jae, et al. “Interaction of Drosophila Melanogaster Prohormone Convertase
2 and 7B2: Insect Cell Specific Processing and Secretion.” Journal of Biological
Chemistry, vol. 275, no. 23, American Society for Biochemistry and Molecular
Biology, 2000, pp. 17886–93, doi:10.1074/jbc.M000032200 .'
short: J. Hwang, D.E. Siekhaus, R. Fuller, P. Taghert, I. Lindberg, Journal of Biological
Chemistry 275 (2000) 17886–17893.
date_created: 2018-12-11T12:01:40Z
date_published: 2000-06-09T00:00:00Z
date_updated: 2023-05-03T08:47:13Z
day: '09'
doi: '10.1074/jbc.M000032200 '
extern: '1'
external_id:
pmid:
- '10749852'
intvolume: ' 275'
issue: '23'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925819833215?via%3Dihub
month: '06'
oa: 1
oa_version: Published Version
page: 17886 - 17893
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '3546'
quality_controlled: '1'
scopus_import: '1'
status: public
title: 'Interaction of Drosophila melanogaster prohormone convertase 2 and 7B2: Insect
cell specific processing and secretion'
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 275
year: '2000'
...
---
_id: '4179'
abstract:
- lang: eng
text: Neurotrophin-3 (NT-3) is a member of the neurotrophin gene family and is highly
expressed in the developing rat cerebellum. Here we show that brain-derived neurotrophic
factor (BDNF) increased by approximately 10-fold the NT-3 mRNA levels in cultured
cerebellar granule neurons isolated from postnatal rats, whereas nerve growth
factor (NGF) and NT-3 itself had no effect. The effect of BDNF was additive to
that of triiodothyronine (T3), which also increased NT-3 mRNA in these neurons.
The drug K252a inhibited the BDNF-mediated stimulation of NT-3 expression, suggesting
an involvement of trkB receptors. Nuclear run-on experiments showed that BDNF
enhanced NT-3 transcription, whereas the stability of NT-3 mRNA remained unchanged.
The data presented are the first demonstration that one neurotrophin regulates
the expression of another and provide evidence that NT-3 production in granule
neurons is regulated by both BDNF and T3.
acknowledgement: We thank Dorothea Stratmann and Karin Angermayer for skillful technical
assistance.
article_processing_charge: No
article_type: original
author:
- first_name: Axel
full_name: Leingärtner, Axel
last_name: Leingärtner
- first_name: Carl-Philipp J
full_name: Heisenberg, Carl-Philipp J
id: 39427864-F248-11E8-B48F-1D18A9856A87
last_name: Heisenberg
orcid: 0000-0002-0912-4566
- first_name: Roland
full_name: Kolbeck, Roland
last_name: Kolbeck
- first_name: Hans
full_name: Thoenen, Hans
last_name: Thoenen
- first_name: Dan
full_name: Lindholm, Dan
last_name: Lindholm
citation:
ama: Leingärtner A, Heisenberg C-PJ, Kolbeck R, Thoenen H, Lindholm D. Brain-derived
neurotrophic factor increases neurotrophin-3 expression in cerebellar granule
neurons. Journal of Biological Chemistry. 1994;269(2):828-830. doi:10.1016/s0021-9258(17)42186-7
apa: Leingärtner, A., Heisenberg, C.-P. J., Kolbeck, R., Thoenen, H., & Lindholm,
D. (1994). Brain-derived neurotrophic factor increases neurotrophin-3 expression
in cerebellar granule neurons. Journal of Biological Chemistry. American
Society for Biochemistry and Molecular Biology. https://doi.org/10.1016/s0021-9258(17)42186-7
chicago: Leingärtner, Axel, Carl-Philipp J Heisenberg, Roland Kolbeck, Hans Thoenen,
and Dan Lindholm. “Brain-Derived Neurotrophic Factor Increases Neurotrophin-3
Expression in Cerebellar Granule Neurons.” Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology, 1994. https://doi.org/10.1016/s0021-9258(17)42186-7.
ieee: A. Leingärtner, C.-P. J. Heisenberg, R. Kolbeck, H. Thoenen, and D. Lindholm,
“Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar
granule neurons,” Journal of Biological Chemistry, vol. 269, no. 2. American
Society for Biochemistry and Molecular Biology, pp. 828–830, 1994.
ista: Leingärtner A, Heisenberg C-PJ, Kolbeck R, Thoenen H, Lindholm D. 1994. Brain-derived
neurotrophic factor increases neurotrophin-3 expression in cerebellar granule
neurons. Journal of Biological Chemistry. 269(2), 828–830.
mla: Leingärtner, Axel, et al. “Brain-Derived Neurotrophic Factor Increases Neurotrophin-3
Expression in Cerebellar Granule Neurons.” Journal of Biological Chemistry,
vol. 269, no. 2, American Society for Biochemistry and Molecular Biology, 1994,
pp. 828–30, doi:10.1016/s0021-9258(17)42186-7.
short: A. Leingärtner, C.-P.J. Heisenberg, R. Kolbeck, H. Thoenen, D. Lindholm,
Journal of Biological Chemistry 269 (1994) 828–830.
date_created: 2018-12-11T12:07:25Z
date_published: 1994-01-14T00:00:00Z
date_updated: 2022-06-02T10:23:48Z
day: '14'
doi: 10.1016/s0021-9258(17)42186-7
extern: '1'
intvolume: ' 269'
issue: '2'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925817421867?via%3Dihub
month: '01'
oa: 1
oa_version: None
page: 828 - 830
publication: Journal of Biological Chemistry
publication_identifier:
eissn:
- 1083-351X
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '1941'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Brain-derived neurotrophic factor increases neurotrophin-3 expression in cerebellar
granule neurons
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 269
year: '1994'
...
---
_id: '2536'
abstract:
- lang: eng
text: A cDNA clone for a new metabotropic glutamate receptor, termed mGluR6, was
isolated from a rat retinal cDNA library by cross-hybridization with the previously
isolated cDNA clone for a metabotropic glutamate receptor. The cloned mGluR6 subtype
consists of 871 amino acid residues and exhibits a structural architecture common
to the metabotropic receptor family, possessing a large extracellular domain preceding
the seven putative membrane-spanning domains. mGluR6 shows the highest sequence
similarity to mGluR4 among the metabotropic receptor subtypes and inhibits the
forskolin- stimulated cyclic AMP accumulation in Chinese hamster ovary cells transfected
with the cloned cDNA. mGluR6 potently reacts with L-2-amino-4- phosphonobutyrate
(L-AP4) and L-serine-O-phosphate, and the potencies of these compounds are one
order of magnitude greater than that of L-glutamate. Blot and in situ hybridization
analyses indicated that mGluR6 mRNA is restrictedly expressed in the inner nuclear
layer of the retina where ON- bipolar cells are distributed. The metabotropic
receptor that responds strongly to L-AP4 and L-serine-O-phosphate in ON-bipolar
cells is known to mediate glutamate synaptic transmission between photoreceptor
cells and ON- bipolar cells. On the basis of the agonist selectivity of mGluR6
and its specific expression in retinal cells, the physiological role of this receptor
subtype in the visual system is discussed.
acknowledgement: "This work was supported in part by research grants from the Ministry
of Education, Science and Culture of Japan, the Ministry of Health and Welfare,
the Yamanouchi Foundation for Research on Metabolic Disorders, the Uehara Memorial
Foundation, and the Inamori Foundation. The costs of publication of this article
were defrayed in part by the payment of page charges. This article must therefore
be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely
to indicate this fact. \r\n\r\nWe are grateful to Akira Uesugi for photographic
assistance."
article_processing_charge: No
article_type: original
author:
- first_name: Yoshiaki
full_name: Nakajima, Yoshiaki
last_name: Nakajima
- first_name: Hideki
full_name: Iwakabe, Hideki
last_name: Iwakabe
- first_name: Chihiro
full_name: Akazawa, Chihiro
last_name: Akazawa
- first_name: Hiroyuki
full_name: Nawa, Hiroyuki
last_name: Nawa
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: Noboru
full_name: Mizuno, Noboru
last_name: Mizuno
- first_name: Shigetada
full_name: Nakanishi, Shigetada
last_name: Nakanishi
citation:
ama: Nakajima Y, Iwakabe H, Akazawa C, et al. Molecular characterization of a novel
retinal metabotropic glutamate receptor mGluR6 with a high agonist selectivity
for L-2-amino-4- phosphonobutyrate. Journal of Biological Chemistry. 1993;268(16):11868-11873.
doi:10.1016/S0021-9258(19)50280-0
apa: Nakajima, Y., Iwakabe, H., Akazawa, C., Nawa, H., Shigemoto, R., Mizuno, N.,
& Nakanishi, S. (1993). Molecular characterization of a novel retinal metabotropic
glutamate receptor mGluR6 with a high agonist selectivity for L-2-amino-4- phosphonobutyrate.
Journal of Biological Chemistry. American Society for Biochemistry and
Molecular Biology. https://doi.org/10.1016/S0021-9258(19)50280-0
chicago: Nakajima, Yoshiaki, Hideki Iwakabe, Chihiro Akazawa, Hiroyuki Nawa, Ryuichi
Shigemoto, Noboru Mizuno, and Shigetada Nakanishi. “Molecular Characterization
of a Novel Retinal Metabotropic Glutamate Receptor MGluR6 with a High Agonist
Selectivity for L-2-Amino-4- Phosphonobutyrate.” Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology, 1993. https://doi.org/10.1016/S0021-9258(19)50280-0.
ieee: Y. Nakajima et al., “Molecular characterization of a novel retinal
metabotropic glutamate receptor mGluR6 with a high agonist selectivity for L-2-amino-4-
phosphonobutyrate,” Journal of Biological Chemistry, vol. 268, no. 16.
American Society for Biochemistry and Molecular Biology, pp. 11868–11873, 1993.
ista: Nakajima Y, Iwakabe H, Akazawa C, Nawa H, Shigemoto R, Mizuno N, Nakanishi
S. 1993. Molecular characterization of a novel retinal metabotropic glutamate
receptor mGluR6 with a high agonist selectivity for L-2-amino-4- phosphonobutyrate.
Journal of Biological Chemistry. 268(16), 11868–11873.
mla: Nakajima, Yoshiaki, et al. “Molecular Characterization of a Novel Retinal Metabotropic
Glutamate Receptor MGluR6 with a High Agonist Selectivity for L-2-Amino-4- Phosphonobutyrate.”
Journal of Biological Chemistry, vol. 268, no. 16, American Society for
Biochemistry and Molecular Biology, 1993, pp. 11868–73, doi:10.1016/S0021-9258(19)50280-0.
short: Y. Nakajima, H. Iwakabe, C. Akazawa, H. Nawa, R. Shigemoto, N. Mizuno, S.
Nakanishi, Journal of Biological Chemistry 268 (1993) 11868–11873.
date_created: 2018-12-11T11:58:15Z
date_published: 1993-06-05T00:00:00Z
date_updated: 2022-04-26T06:56:15Z
day: '05'
doi: 10.1016/S0021-9258(19)50280-0
extern: '1'
external_id:
pmid:
- '8389366'
intvolume: ' 268'
issue: '16'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.1016/S0021-9258(19)50280-0
month: '06'
oa: 1
oa_version: Published Version
page: 11868 - 11873
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4362'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Molecular characterization of a novel retinal metabotropic glutamate receptor
mGluR6 with a high agonist selectivity for L-2-amino-4- phosphonobutyrate
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 268
year: '1993'
...
---
_id: '2539'
abstract:
- lang: eng
text: cDNA clones for four different N-methyl-D-aspartate (NMDA) receptor subunits
(NMDAR2A-NMDAR2D) were isolated through polymerase chain reactions followed by
molecular screening of a rat brain cDNA library. These subunits are only about
15% identical with the key subunit of the NMDA receptor (NMDAR1) but are highly
homologous (~50% homology) with one another. They also commonly possess large
hydrophilic domains at both amino- and carboxyl- terminal sides of the four putative
transmembrane segments. NMDAR2A and NMDAR2C expressed individually in Xenopus
oocytes showed no electrophysiological response to agonists. However, these subunits
in combined expression with NMDAR1 markedly potentiated the NMDAR1 activity and
produced functional variability in the affinity of agonists, the effectiveness
of antagonists, and the sensitivity to Mg2+ blockade. Thus, NMDAR1 is essential
for the function of the NMDA receptor, and multiple NMDAR2 subunits potentiate
and differentiate the function of the NMDA receptor by forming different heteromeric
configurations with NMDAR1. Northern blotting and in situ hybridization analyses
revealed that the expressions of individual mRNAs for the NMDAR2 subunits overlap
in some brain regions but are also specialized in many other regions. This investigation
demonstrates the anatomical and functional differences of the NMDAR2 subunits,
which provide the molecular basis for the functional diversity of the NMDA receptor.
acknowledgement: This work was supported in part by research grants from the Ministry
of Education, Science, and Culture of Japan, the Ministry of Health and Welfare
of Japan, the Senri Life Science Foundation, and Yamanouchi Foundation for Research
on Metabolic Disorders. The costs of publication of this article were defrayed in
part by the payment of page charges. This article must therefore be hereby marked
“aduertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this
fact.
article_processing_charge: No
article_type: original
author:
- first_name: Takahiro
full_name: Ishii, Takahiro
last_name: Ishii
- first_name: Koki
full_name: Moriyoshi, Koki
last_name: Moriyoshi
- first_name: Hidemitsu
full_name: Sugihara, Hidemitsu
last_name: Sugihara
- first_name: Kazuhir
full_name: Sakurada, Kazuhir
last_name: Sakurada
- first_name: Hiroshi
full_name: Kadotani, Hiroshi
last_name: Kadotani
- first_name: Mineto
full_name: Yokoi, Mineto
last_name: Yokoi
- first_name: Chihiro
full_name: Akazawa, Chihiro
last_name: Akazawa
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: Noboru
full_name: Mizuno, Noboru
last_name: Mizuno
- first_name: Masayuki
full_name: Masu, Masayuki
last_name: Masu
- first_name: Shigetada
full_name: Nakanishi, Shigetada
last_name: Nakanishi
citation:
ama: Ishii T, Moriyoshi K, Sugihara H, et al. Molecular characterization of the
family of the N-methyl-D-aspartate receptor subunits. Journal of Biological
Chemistry. 1993;268(4):2836-2843. doi:10.1016/s0021-9258(18)53849-7
apa: Ishii, T., Moriyoshi, K., Sugihara, H., Sakurada, K., Kadotani, H., Yokoi,
M., … Nakanishi, S. (1993). Molecular characterization of the family of the N-methyl-D-aspartate
receptor subunits. Journal of Biological Chemistry. American Society for
Biochemistry and Molecular Biology. https://doi.org/10.1016/s0021-9258(18)53849-7
chicago: Ishii, Takahiro, Koki Moriyoshi, Hidemitsu Sugihara, Kazuhir Sakurada,
Hiroshi Kadotani, Mineto Yokoi, Chihiro Akazawa, et al. “Molecular Characterization
of the Family of the N-Methyl-D-Aspartate Receptor Subunits.” Journal of Biological
Chemistry. American Society for Biochemistry and Molecular Biology, 1993.
https://doi.org/10.1016/s0021-9258(18)53849-7
.
ieee: T. Ishii et al., “Molecular characterization of the family of the N-methyl-D-aspartate
receptor subunits,” Journal of Biological Chemistry, vol. 268, no. 4. American
Society for Biochemistry and Molecular Biology, pp. 2836–2843, 1993.
ista: Ishii T, Moriyoshi K, Sugihara H, Sakurada K, Kadotani H, Yokoi M, Akazawa
C, Shigemoto R, Mizuno N, Masu M, Nakanishi S. 1993. Molecular characterization
of the family of the N-methyl-D-aspartate receptor subunits. Journal of Biological
Chemistry. 268(4), 2836–2843.
mla: Ishii, Takahiro, et al. “Molecular Characterization of the Family of the N-Methyl-D-Aspartate
Receptor Subunits.” Journal of Biological Chemistry, vol. 268, no. 4, American
Society for Biochemistry and Molecular Biology, 1993, pp. 2836–43, doi:10.1016/s0021-9258(18)53849-7 .
short: T. Ishii, K. Moriyoshi, H. Sugihara, K. Sakurada, H. Kadotani, M. Yokoi,
C. Akazawa, R. Shigemoto, N. Mizuno, M. Masu, S. Nakanishi, Journal of Biological
Chemistry 268 (1993) 2836–2843.
date_created: 2018-12-11T11:58:16Z
date_published: 1993-02-05T00:00:00Z
date_updated: 2022-03-31T14:29:17Z
day: '05'
doi: '10.1016/s0021-9258(18)53849-7 '
extern: '1'
external_id:
pmid:
- '8428958'
intvolume: ' 268'
issue: '4'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.jbc.org/article/S0021-9258(18)53849-7/fulltext
month: '02'
oa: 1
oa_version: Published Version
page: 2836 - 2843
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4360'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Molecular characterization of the family of the N-methyl-D-aspartate receptor
subunits
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 268
year: '1993'
...
---
_id: '2533'
abstract:
- lang: eng
text: A cDNA clone for a new metabotropic glutamate receptor, mGluR5, was isolated
through polymerase chain reaction-mediated DNA amplification by using primer sequences
conserved among the metabotropic glutamate receptor (mGluR) family and by the
subsequent screening of a rat brain cDNA library. The cloned receptor consists
of 1171 amino acid residues and exhibits a structural architecture common to the
mGluR family, possessing a large extracellular domain preceding the seven putative
membrane-spanning segments. mGluR5 shows the highest sequence similarity to mGluR1
among the mGluR members and is coupled to the stimulation of phosphatidylinositol
hydrolysis/ Ca2+ signal transduction in Chinese hamster ovary cells transfected
with the cloned cDNA. This receptor also resembles mGluR1 in its agonist selectivity
and antagonist responses; the potency rank order of agonists for mGluR5 was determined
to be quisqualate > L-glutamate ≥ ibotenate > trans-1-aminocyclopentane-1,3-dicarboxylate.
Blot and in situ hybridization analyses indicated that mGluR5 mRNA is widely distributed
in neuronal cells of the central nervous system and is expressed differently from
mGluR1 mRNA in many brain regions. This investigation thus demonstrates that there
is an additional mGluR subtype which closely resembles mGluR1 in its signal transduction
and pharmacological properties and is expressed in specialized neuronal cells
in the central nervous system.
acknowledgement: We are grateful to Seiji Ito for help of Ca2+ measurements and Akira
Uesugi for photographic assistance.
article_processing_charge: No
article_type: original
author:
- first_name: Takaaki
full_name: Abe, Takaaki
last_name: Abe
- first_name: Hidemitsu
full_name: Sugihara, Hidemitsu
last_name: Sugihara
- first_name: Hiroyuki
full_name: Nawa, Hiroyuki
last_name: Nawa
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: Noboru
full_name: Mizuno, Noboru
last_name: Mizuno
- first_name: Shigetada
full_name: Nakanishi, Shigetada
last_name: Nakanishi
citation:
ama: Abe T, Sugihara H, Nawa H, Shigemoto R, Mizuno N, Nakanishi S. Molecular characterization
of a novel metabotropic glutamate receptor mGluR5 coupled to inositol phosphate/Ca2+
signal transduction. Journal of Biological Chemistry. 1992;267(19):13361-13368.
doi:10.1016/S0021-9258(18)42219-3
apa: Abe, T., Sugihara, H., Nawa, H., Shigemoto, R., Mizuno, N., & Nakanishi,
S. (1992). Molecular characterization of a novel metabotropic glutamate receptor
mGluR5 coupled to inositol phosphate/Ca2+ signal transduction. Journal of Biological
Chemistry. American Society for Biochemistry and Molecular Biology. https://doi.org/10.1016/S0021-9258(18)42219-3
chicago: Abe, Takaaki, Hidemitsu Sugihara, Hiroyuki Nawa, Ryuichi Shigemoto, Noboru
Mizuno, and Shigetada Nakanishi. “Molecular Characterization of a Novel Metabotropic
Glutamate Receptor MGluR5 Coupled to Inositol Phosphate/Ca2+ Signal Transduction.”
Journal of Biological Chemistry. American Society for Biochemistry and
Molecular Biology, 1992. https://doi.org/10.1016/S0021-9258(18)42219-3.
ieee: T. Abe, H. Sugihara, H. Nawa, R. Shigemoto, N. Mizuno, and S. Nakanishi, “Molecular
characterization of a novel metabotropic glutamate receptor mGluR5 coupled to
inositol phosphate/Ca2+ signal transduction,” Journal of Biological Chemistry,
vol. 267, no. 19. American Society for Biochemistry and Molecular Biology, pp.
13361–13368, 1992.
ista: Abe T, Sugihara H, Nawa H, Shigemoto R, Mizuno N, Nakanishi S. 1992. Molecular
characterization of a novel metabotropic glutamate receptor mGluR5 coupled to
inositol phosphate/Ca2+ signal transduction. Journal of Biological Chemistry.
267(19), 13361–13368.
mla: Abe, Takaaki, et al. “Molecular Characterization of a Novel Metabotropic Glutamate
Receptor MGluR5 Coupled to Inositol Phosphate/Ca2+ Signal Transduction.” Journal
of Biological Chemistry, vol. 267, no. 19, American Society for Biochemistry
and Molecular Biology, 1992, pp. 13361–68, doi:10.1016/S0021-9258(18)42219-3.
short: T. Abe, H. Sugihara, H. Nawa, R. Shigemoto, N. Mizuno, S. Nakanishi, Journal
of Biological Chemistry 267 (1992) 13361–13368.
date_created: 2018-12-11T11:58:14Z
date_published: 1992-07-05T00:00:00Z
date_updated: 2022-03-17T15:08:29Z
day: '05'
doi: 10.1016/S0021-9258(18)42219-3
extern: '1'
external_id:
pmid:
- '1320017'
intvolume: ' 267'
issue: '19'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925818422193
month: '07'
oa: 1
oa_version: Published Version
page: 13361 - 13368
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4366'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Molecular characterization of a novel metabotropic glutamate receptor mGluR5
coupled to inositol phosphate/Ca2+ signal transduction
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 267
year: '1992'
...
---
_id: '2480'
abstract:
- lang: eng
text: Functional cDNA clones for rat neuromedin K receptor were isolated from a
rat brain cDNA library by cross-hybridization with the bovine substance K recepor
cDNA. Injection of the mRNA synthesized in vitro from the cloned cDNA into Xenopus
oocytes elicited electrophysiological responses to tachykinins, with the most
potent sensitivity being to neuromedin K. Ligand-binding displacement in membranes
of mammalian COS cells transfected with the cDNA indicated the rank order of affinity
of the receptor to tachykinins; neuromedin K > substance K > substance P.
The hybridization analysis showed that the neuromedin K receptor mRNA is expressed
in both the brain and the peripheral tissues at different levels. The rat neuromedin
K receptor consists of 452 amino acid residues and belongs to the family of G
protein-coupled receptors, which are thought to have seven transmembrane domains.
The sequence comparison of the rat neuromedin K, substance P, and substance K
receptors revealed that these receptors are highly conserved in the seven transmembrane
domains and the cytoplasmic sides of the receptors. They also show some structural
characteristics, including the common presence of histidine residues in transmembrane
segments V and VI and the difference in the numbers and distributions of serine
and threonine residues as possible phosphorylation sites in the cytoplasmic regions.
This paper thus presents the first comprehensive analysis of the molecular nature
of the multiple peptide receptors that exhibit similar but pharmacologically distinguishable
activities.
acknowledgement: This work was supported in part by research grants from the Ministry
Education, Science and Culture of Japan; the Institute of Physical and Chemical
Research; and the Science and Technology Agency of Japan. The costs of publication
of this article were defrayed in part by the payment of page charges. This article
must therefore be hereby marked “advertisement” in accordance with 18 USC. Section
1734 solely to indicate this fact.
article_processing_charge: No
article_type: original
author:
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: Yoshifumi
full_name: Yokota, Yoshifumi
last_name: Yokota
- first_name: Kunihiro
full_name: Tsuchida, Kunihiro
last_name: Tsuchida
- first_name: Shigetada
full_name: Nakanishi, Shigetada
last_name: Nakanishi
citation:
ama: Shigemoto R, Yokota Y, Tsuchida K, Nakanishi S. Cloning and expression of a
rat neuromedin K receptor cDNA. Journal of Biological Chemistry. 1990;265(2):623-628.
doi:10.1016/s0021-9258(19)40095-1
apa: Shigemoto, R., Yokota, Y., Tsuchida, K., & Nakanishi, S. (1990). Cloning
and expression of a rat neuromedin K receptor cDNA. Journal of Biological Chemistry.
American Society for Biochemistry and Molecular Biology. https://doi.org/10.1016/s0021-9258(19)40095-1
chicago: Shigemoto, Ryuichi, Yoshifumi Yokota, Kunihiro Tsuchida, and Shigetada
Nakanishi. “Cloning and Expression of a Rat Neuromedin K Receptor CDNA.” Journal
of Biological Chemistry. American Society for Biochemistry and Molecular Biology,
1990. https://doi.org/10.1016/s0021-9258(19)40095-1
.
ieee: R. Shigemoto, Y. Yokota, K. Tsuchida, and S. Nakanishi, “Cloning and expression
of a rat neuromedin K receptor cDNA,” Journal of Biological Chemistry,
vol. 265, no. 2. American Society for Biochemistry and Molecular Biology, pp.
623–628, 1990.
ista: Shigemoto R, Yokota Y, Tsuchida K, Nakanishi S. 1990. Cloning and expression
of a rat neuromedin K receptor cDNA. Journal of Biological Chemistry. 265(2),
623–628.
mla: Shigemoto, Ryuichi, et al. “Cloning and Expression of a Rat Neuromedin K Receptor
CDNA.” Journal of Biological Chemistry, vol. 265, no. 2, American Society
for Biochemistry and Molecular Biology, 1990, pp. 623–28, doi:10.1016/s0021-9258(19)40095-1 .
short: R. Shigemoto, Y. Yokota, K. Tsuchida, S. Nakanishi, Journal of Biological
Chemistry 265 (1990) 623–628.
date_created: 2018-12-11T11:57:55Z
date_published: 1990-01-15T00:00:00Z
date_updated: 2022-02-24T11:07:05Z
day: '15'
doi: '10.1016/s0021-9258(19)40095-1 '
extern: '1'
external_id:
pmid:
- '2153106 '
intvolume: ' 265'
issue: '2'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925819400951
month: '01'
oa: 1
oa_version: Published Version
page: 623 - 628
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
eissn:
- 1083-351X
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4421'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Cloning and expression of a rat neuromedin K receptor cDNA
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 265
year: '1990'
...
---
_id: '2525'
abstract:
- lang: eng
text: This paper describes the amino acid sequence of the rat substance P receptor
and its comparison with that of the rat substance K receptor on the basis of molecular
cloning and sequence analysis. From a rat brain cDNA library constructed with
an RNA expression vector, we identified a cDNA mixture containing a functional
substance P receptor cDNA by examining electrophysiologically a receptor expression
following injection of the mRNAs synthesized in vitro into Xenopus oocytes. A
receptor cDNA clone was then isolated by cross-hybridization with the bovine substance
K receptor DNA. The clone was confirmed by selective binding of substance P to
the cloned receptor expressed in mammalian COS cells. The deduced amino acid sequence
(407 amino acid residues) possesses seven putative membrane spanning domains and
shows a sequence similarity to the members of G-protein-coupled receptors. The
rat substance P and substance K receptor are very similar in both size and amino
acid sequences, particularly in the putative transmembrane similarity is in marked
contrast to the sequence divergence in the amino- and carboxyl-terminal regions
and the third cytoplasmic loop. The observed sequence similarytity and divergence
would thus contribute to the expression of similar but pharmacological regions
and the first and second cytoplasmic loops. This distinguishable activities of
the two tachykinin receptors.
acknowledgement: 'This work was supported in part by research grants from the Ministry
of Education, Science and Culture of Japan, the Institute of Physical and Chemical
Research, and the Science and Technology Agency of Japan. The costs of publication
of this article were defrayed in part by the payment of page charges. This article
must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section
1734 solely to indicate this fact. '
article_processing_charge: No
article_type: original
author:
- first_name: Yoshifumi
full_name: Yokota, Yoshifumi
last_name: Yokota
- first_name: Yoshiki
full_name: Sasai, Yoshiki
last_name: Sasai
- first_name: Kohichi
full_name: Tanaka, Kohichi
last_name: Tanaka
- first_name: Tsutomu
full_name: Fujiwara, Tsutomu
last_name: Fujiwara
- first_name: Kunihiro
full_name: Tsuchida, Kunihiro
last_name: Tsuchida
- first_name: Ryuichi
full_name: Shigemoto, Ryuichi
id: 499F3ABC-F248-11E8-B48F-1D18A9856A87
last_name: Shigemoto
orcid: 0000-0001-8761-9444
- first_name: Akira
full_name: Kakizuka, Akira
last_name: Kakizuka
- first_name: Hiroaki
full_name: Ohkubo, Hiroaki
last_name: Ohkubo
- first_name: Shigetada
full_name: Nakanishi, Shigetada
last_name: Nakanishi
citation:
ama: Yokota Y, Sasai Y, Tanaka K, et al. Molecular characterization of a functional
cDNA for rat substance P receptor. Journal of Biological Chemistry. 1989;264(30):17649-17652.
doi:doi.org/10.1016/S0021-9258(19)84619-7
apa: Yokota, Y., Sasai, Y., Tanaka, K., Fujiwara, T., Tsuchida, K., Shigemoto, R.,
… Nakanishi, S. (1989). Molecular characterization of a functional cDNA for rat
substance P receptor. Journal of Biological Chemistry. American Society
for Biochemistry and Molecular Biology. https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7
chicago: Yokota, Yoshifumi, Yoshiki Sasai, Kohichi Tanaka, Tsutomu Fujiwara, Kunihiro
Tsuchida, Ryuichi Shigemoto, Akira Kakizuka, Hiroaki Ohkubo, and Shigetada Nakanishi.
“Molecular Characterization of a Functional CDNA for Rat Substance P Receptor.”
Journal of Biological Chemistry. American Society for Biochemistry and
Molecular Biology, 1989. https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7.
ieee: Y. Yokota et al., “Molecular characterization of a functional cDNA
for rat substance P receptor,” Journal of Biological Chemistry, vol. 264,
no. 30. American Society for Biochemistry and Molecular Biology, pp. 17649–17652,
1989.
ista: Yokota Y, Sasai Y, Tanaka K, Fujiwara T, Tsuchida K, Shigemoto R, Kakizuka
A, Ohkubo H, Nakanishi S. 1989. Molecular characterization of a functional cDNA
for rat substance P receptor. Journal of Biological Chemistry. 264(30), 17649–17652.
mla: Yokota, Yoshifumi, et al. “Molecular Characterization of a Functional CDNA
for Rat Substance P Receptor.” Journal of Biological Chemistry, vol. 264,
no. 30, American Society for Biochemistry and Molecular Biology, 1989, pp. 17649–52,
doi:doi.org/10.1016/S0021-9258(19)84619-7.
short: Y. Yokota, Y. Sasai, K. Tanaka, T. Fujiwara, K. Tsuchida, R. Shigemoto, A.
Kakizuka, H. Ohkubo, S. Nakanishi, Journal of Biological Chemistry 264 (1989)
17649–17652.
date_created: 2018-12-11T11:58:11Z
date_published: 1989-10-25T00:00:00Z
date_updated: 2022-02-15T09:29:36Z
day: '25'
doi: doi.org/10.1016/S0021-9258(19)84619-7
extern: '1'
external_id:
pmid:
- '2478537'
intvolume: ' 264'
issue: '30'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.sciencedirect.com/science/article/pii/S0021925819846197
month: '10'
oa: 1
oa_version: Published Version
page: 17649 - 17652
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
eissn:
- 1083-351X
issn:
- 0021-9258
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '4374'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Molecular characterization of a functional cDNA for rat substance P receptor
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 264
year: '1989'
...