---
_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. <i>Journal of Biological Chemistry</i>. 2022;298(9). doi:<a
    href="https://doi.org/10.1016/j.jbc.2022.102343">10.1016/j.jbc.2022.102343</a>
  apa: Artan, M., Hartl, M., Chen, W., &#38; de Bono, M. (2022). Depletion of endogenously
    biotinylated carboxylases enhances the sensitivity of TurboID-mediated proximity
    labeling in Caenorhabditis elegans. <i>Journal of Biological Chemistry</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.jbc.2022.102343">https://doi.org/10.1016/j.jbc.2022.102343</a>
  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.” <i>Journal of Biological Chemistry</i>.
    Elsevier, 2022. <a href="https://doi.org/10.1016/j.jbc.2022.102343">https://doi.org/10.1016/j.jbc.2022.102343</a>.
  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,” <i>Journal of Biological Chemistry</i>, 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.”
    <i>Journal of Biological Chemistry</i>, vol. 298, no. 9, 102343, Elsevier, 2022,
    doi:<a href="https://doi.org/10.1016/j.jbc.2022.102343">10.1016/j.jbc.2022.102343</a>.
  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
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. <i>Journal of Biological
    Chemistry</i>. 2021;297(3). doi:<a href="https://doi.org/10.1016/J.JBC.2021.101094">10.1016/J.JBC.2021.101094</a>
  apa: Artan, M., Barratt, S., Flynn, S. M., Begum, F., Skehel, M., Nicolas, A., &#38;
    de Bono, M. (2021). Interactome analysis of Caenorhabditis elegans synapses by
    TurboID-based proximity labeling. <i>Journal of Biological Chemistry</i>. Elsevier.
    <a href="https://doi.org/10.1016/J.JBC.2021.101094">https://doi.org/10.1016/J.JBC.2021.101094</a>
  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.” <i>Journal of Biological Chemistry</i>.
    Elsevier, 2021. <a href="https://doi.org/10.1016/J.JBC.2021.101094">https://doi.org/10.1016/J.JBC.2021.101094</a>.
  ieee: M. Artan <i>et al.</i>, “Interactome analysis of Caenorhabditis elegans synapses
    by TurboID-based proximity labeling,” <i>Journal of Biological Chemistry</i>,
    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.” <i>Journal of Biological Chemistry</i>,
    vol. 297, no. 3, 101094, Elsevier, 2021, doi:<a href="https://doi.org/10.1016/J.JBC.2021.101094">10.1016/J.JBC.2021.101094</a>.
  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: '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. <i>Journal of Biological Chemistry</i>. 1994;269(2):828-830. doi:<a href="https://doi.org/10.1016/s0021-9258(17)42186-7">10.1016/s0021-9258(17)42186-7</a>
  apa: Leingärtner, A., Heisenberg, C.-P. J., Kolbeck, R., Thoenen, H., &#38; Lindholm,
    D. (1994). Brain-derived neurotrophic factor increases neurotrophin-3 expression
    in cerebellar granule neurons. <i>Journal of Biological Chemistry</i>. American
    Society for Biochemistry and Molecular Biology. <a href="https://doi.org/10.1016/s0021-9258(17)42186-7">https://doi.org/10.1016/s0021-9258(17)42186-7</a>
  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.” <i>Journal of Biological Chemistry</i>.
    American Society for Biochemistry and Molecular Biology, 1994. <a href="https://doi.org/10.1016/s0021-9258(17)42186-7">https://doi.org/10.1016/s0021-9258(17)42186-7</a>.
  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,” <i>Journal of Biological Chemistry</i>, 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.” <i>Journal of Biological Chemistry</i>,
    vol. 269, no. 2, American Society for Biochemistry and Molecular Biology, 1994,
    pp. 828–30, doi:<a href="https://doi.org/10.1016/s0021-9258(17)42186-7">10.1016/s0021-9258(17)42186-7</a>.
  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: '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 &gt; substance K &gt; 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. <i>Journal of Biological Chemistry</i>. 1990;265(2):623-628.
    doi:<a href="https://doi.org/10.1016/s0021-9258(19)40095-1 ">10.1016/s0021-9258(19)40095-1
    </a>
  apa: Shigemoto, R., Yokota, Y., Tsuchida, K., &#38; Nakanishi, S. (1990). Cloning
    and expression of a rat neuromedin K receptor cDNA. <i>Journal of Biological Chemistry</i>.
    American Society for Biochemistry and Molecular Biology. <a href="https://doi.org/10.1016/s0021-9258(19)40095-1
    ">https://doi.org/10.1016/s0021-9258(19)40095-1 </a>
  chicago: Shigemoto, Ryuichi, Yoshifumi Yokota, Kunihiro Tsuchida, and Shigetada
    Nakanishi. “Cloning and Expression of a Rat Neuromedin K Receptor CDNA.” <i>Journal
    of Biological Chemistry</i>. American Society for Biochemistry and Molecular Biology,
    1990. <a href="https://doi.org/10.1016/s0021-9258(19)40095-1 ">https://doi.org/10.1016/s0021-9258(19)40095-1
    </a>.
  ieee: R. Shigemoto, Y. Yokota, K. Tsuchida, and S. Nakanishi, “Cloning and expression
    of a rat neuromedin K receptor cDNA,” <i>Journal of Biological Chemistry</i>,
    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.” <i>Journal of Biological Chemistry</i>, vol. 265, no. 2, American Society
    for Biochemistry and Molecular Biology, 1990, pp. 623–28, doi:<a href="https://doi.org/10.1016/s0021-9258(19)40095-1
    ">10.1016/s0021-9258(19)40095-1 </a>.
  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. <i>Journal of Biological Chemistry</i>. 1989;264(30):17649-17652.
    doi:<a href="https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7">doi.org/10.1016/S0021-9258(19)84619-7</a>
  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. <i>Journal of Biological Chemistry</i>. American Society
    for Biochemistry and Molecular Biology. <a href="https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7">https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7</a>
  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.”
    <i>Journal of Biological Chemistry</i>. American Society for Biochemistry and
    Molecular Biology, 1989. <a href="https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7">https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7</a>.
  ieee: Y. Yokota <i>et al.</i>, “Molecular characterization of a functional cDNA
    for rat substance P receptor,” <i>Journal of Biological Chemistry</i>, 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.” <i>Journal of Biological Chemistry</i>, vol. 264,
    no. 30, American Society for Biochemistry and Molecular Biology, 1989, pp. 17649–52,
    doi:<a href="https://doi.org/doi.org/10.1016/S0021-9258(19)84619-7">doi.org/10.1016/S0021-9258(19)84619-7</a>.
  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'
...