---
_id: '12163'
abstract:
- lang: eng
text: Small GTPases play essential roles in the organization of eukaryotic cells.
In recent years, it has become clear that their intracellular functions result
from intricate biochemical networks of the GTPase and their regulators that dynamically
bind to a membrane surface. Due to the inherent complexities of their interactions,
however, revealing the underlying mechanisms of action is often difficult to achieve
from in vivo studies. This review summarizes in vitro reconstitution approaches
developed to obtain a better mechanistic understanding of how small GTPase activities
are regulated in space and time.
acknowledgement: The authors acknowledge support from IST Austria and helpful comments
from the anonymous reviewers that helped to improve this manuscript. We apologize
to the authors of primary literature and outstanding research not cited here due
to space restraints.
article_processing_charge: Yes (via OA deal)
article_type: review
author:
- first_name: Martin
full_name: Loose, Martin
id: 462D4284-F248-11E8-B48F-1D18A9856A87
last_name: Loose
orcid: 0000-0001-7309-9724
- first_name: Albert
full_name: Auer, Albert
id: 3018E8C2-F248-11E8-B48F-1D18A9856A87
last_name: Auer
orcid: 0000-0002-3580-2906
- first_name: Gabriel
full_name: Brognara, Gabriel
id: D96FFDA0-A884-11E9-9968-DC26E6697425
last_name: Brognara
- first_name: Hanifatul R
full_name: Budiman, Hanifatul R
id: 55380f95-15b2-11ec-abd3-aff8e230696b
last_name: Budiman
- first_name: Lukasz M
full_name: Kowalski, Lukasz M
id: e3a512e2-4bbe-11eb-a68a-e3857a7844c2
last_name: Kowalski
- first_name: Ivana
full_name: Matijevic, Ivana
id: 83c17ce3-15b2-11ec-abd3-f486545870bd
last_name: Matijevic
citation:
ama: Loose M, Auer A, Brognara G, Budiman HR, Kowalski LM, Matijevic I. In vitro
reconstitution of small GTPase regulation. FEBS Letters. 2023;597(6):762-777.
doi:10.1002/1873-3468.14540
apa: Loose, M., Auer, A., Brognara, G., Budiman, H. R., Kowalski, L. M., & Matijevic,
I. (2023). In vitro reconstitution of small GTPase regulation. FEBS Letters.
Wiley. https://doi.org/10.1002/1873-3468.14540
chicago: Loose, Martin, Albert Auer, Gabriel Brognara, Hanifatul R Budiman, Lukasz
M Kowalski, and Ivana Matijevic. “In Vitro Reconstitution of Small GTPase Regulation.”
FEBS Letters. Wiley, 2023. https://doi.org/10.1002/1873-3468.14540.
ieee: M. Loose, A. Auer, G. Brognara, H. R. Budiman, L. M. Kowalski, and I. Matijevic,
“In vitro reconstitution of small GTPase regulation,” FEBS Letters, vol.
597, no. 6. Wiley, pp. 762–777, 2023.
ista: Loose M, Auer A, Brognara G, Budiman HR, Kowalski LM, Matijevic I. 2023. In
vitro reconstitution of small GTPase regulation. FEBS Letters. 597(6), 762–777.
mla: Loose, Martin, et al. “In Vitro Reconstitution of Small GTPase Regulation.”
FEBS Letters, vol. 597, no. 6, Wiley, 2023, pp. 762–77, doi:10.1002/1873-3468.14540.
short: M. Loose, A. Auer, G. Brognara, H.R. Budiman, L.M. Kowalski, I. Matijevic,
FEBS Letters 597 (2023) 762–777.
date_created: 2023-01-12T12:09:58Z
date_published: 2023-03-01T00:00:00Z
date_updated: 2023-08-16T08:32:29Z
day: '01'
ddc:
- '570'
department:
- _id: MaLo
doi: 10.1002/1873-3468.14540
external_id:
isi:
- '000891573000001'
pmid:
- '36448231'
file:
- access_level: open_access
checksum: 7492244d3f9c5faa1347ef03f6e5bc84
content_type: application/pdf
creator: dernst
date_created: 2023-08-16T08:31:04Z
date_updated: 2023-08-16T08:31:04Z
file_id: '14063'
file_name: 2023_FEBSLetters_Loose.pdf
file_size: 3148143
relation: main_file
success: 1
file_date_updated: 2023-08-16T08:31:04Z
has_accepted_license: '1'
intvolume: ' 597'
isi: 1
issue: '6'
keyword:
- Cell Biology
- Genetics
- Molecular Biology
- Biochemistry
- Structural Biology
- Biophysics
language:
- iso: eng
month: '03'
oa: 1
oa_version: Published Version
page: 762-777
pmid: 1
publication: FEBS Letters
publication_identifier:
eissn:
- 1873-3468
issn:
- 0014-5793
publication_status: published
publisher: Wiley
quality_controlled: '1'
scopus_import: '1'
status: public
title: In vitro reconstitution of small GTPase regulation
tmp:
image: /images/cc_by_nc_nd.png
legal_code_url: https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode
name: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
(CC BY-NC-ND 4.0)
short: CC BY-NC-ND (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 597
year: '2023'
...
---
_id: '1954'
abstract:
- lang: eng
text: "\r\nWe have examined the effects of heat stress on electron transfer in the
thylakoid membrane of an engineered plastid ndh deletion mutant, Δ1, incapable
of performing the Ndh-mediated reduction of the plastoquinone pool in the chloroplast.
Upon heat stress in the dark, the rate of PSII- independent reduction of PSI after
subsequent illumination by far-red light is dramatically enhanced in both Δ1 and
a wild-type control plant (WT). In contrast, in the dark, only the WT shows an
increase in the reduction state of the plastoquinone pool. We conclude that the
heat stress-induced reduction of the intersystem electron transport chain can
be mediated by Ndh- independent pathways in the light but that in the dark the
dominant pathway for reduction of the plastoquinone pool is catalysed by the Ndh
complex. Our results therefore demonstrate a functional role for the Ndh complex
in the dark.\r\n"
acknowledgement: This work was funded by the BBSRC. We would like to thank Professor
Peter Horton (University of Sheffield) for the loan of the ED 800 T unit.
article_processing_charge: No
article_type: original
author:
- first_name: Leonid A
full_name: Sazanov, Leonid A
id: 338D39FE-F248-11E8-B48F-1D18A9856A87
last_name: Sazanov
orcid: 0000-0002-0977-7989
- first_name: Paul
full_name: Burrows, Paul
last_name: Burrows
- first_name: Peter
full_name: Nixon, Peter
last_name: Nixon
citation:
ama: Sazanov LA, Burrows P, Nixon P. The chloroplast Ndh complex mediates the dark
reduction of the plastoquinone pool in response to heat stress in tobacco leaves.
FEBS Letters. 1998;429(1):115-118. doi:10.1016/S0014-5793(98)00573-0
apa: Sazanov, L. A., Burrows, P., & Nixon, P. (1998). The chloroplast Ndh complex
mediates the dark reduction of the plastoquinone pool in response to heat stress
in tobacco leaves. FEBS Letters. Elsevier. https://doi.org/10.1016/S0014-5793(98)00573-0
chicago: Sazanov, Leonid A, Paul Burrows, and Peter Nixon. “The Chloroplast Ndh
Complex Mediates the Dark Reduction of the Plastoquinone Pool in Response to Heat
Stress in Tobacco Leaves.” FEBS Letters. Elsevier, 1998. https://doi.org/10.1016/S0014-5793(98)00573-0.
ieee: L. A. Sazanov, P. Burrows, and P. Nixon, “The chloroplast Ndh complex mediates
the dark reduction of the plastoquinone pool in response to heat stress in tobacco
leaves,” FEBS Letters, vol. 429, no. 1. Elsevier, pp. 115–118, 1998.
ista: Sazanov LA, Burrows P, Nixon P. 1998. The chloroplast Ndh complex mediates
the dark reduction of the plastoquinone pool in response to heat stress in tobacco
leaves. FEBS Letters. 429(1), 115–118.
mla: Sazanov, Leonid A., et al. “The Chloroplast Ndh Complex Mediates the Dark Reduction
of the Plastoquinone Pool in Response to Heat Stress in Tobacco Leaves.” FEBS
Letters, vol. 429, no. 1, Elsevier, 1998, pp. 115–18, doi:10.1016/S0014-5793(98)00573-0.
short: L.A. Sazanov, P. Burrows, P. Nixon, FEBS Letters 429 (1998) 115–118.
date_created: 2018-12-11T11:54:54Z
date_published: 1998-06-05T00:00:00Z
date_updated: 2022-09-01T13:12:15Z
day: '05'
doi: 10.1016/S0014-5793(98)00573-0
extern: '1'
external_id:
pmid:
- '9657394 '
intvolume: ' 429'
issue: '1'
language:
- iso: eng
month: '06'
oa_version: None
page: 115 - 118
pmid: 1
publication: FEBS Letters
publication_identifier:
issn:
- 0014-5793
publication_status: published
publisher: Elsevier
publist_id: '5128'
quality_controlled: '1'
status: public
title: The chloroplast Ndh complex mediates the dark reduction of the plastoquinone
pool in response to heat stress in tobacco leaves
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 429
year: '1998'
...
---
_id: '1949'
abstract:
- lang: eng
text: H+-transhydrogenase (H+-Thase) and NADP-linked isocitrate dehydrogenase (NADP-ICDH)
are very active in animal mitochondria but their physiological function is only
poorly understood. This is especially so in the case of the heart and muscle,
where there are no major consumers of NADPH. We propose here that H+-Thase and
NADP-ICDH have a combined function in the fine regulation of the activity of the
tricarboxylic acid (TCA) cycle, providing enhanced sensitivy to changes in energy
demand. This is achieved through cycling of substrates by NAD-linked ICDH, NADP-linked
ICDH and H+-Thase. It is proposed that NAD-ICDH operates in the forward direction
of the TCA cycle, but NADP-ICDH is driven in reverse by elevated levels of NADPH
resulting from the action of the transmembrane proton electrochemical potential
gradient (Δp) on H+-Thase. This has the effect of increasing the sensitivity to
allosteric modifiers of NAD-ICDH (NADH, ADP, ATP, Ca2+ etc), potentially giving
rise to large changes in the net flux from iso-citrate to α-ketoglutarate. Furthermore,
changes in the level of Δp resulting from changes in the demand for ATP would,
via H+-Thase, shift the redox state of the NADP pool and this, in turn, would
lead to a change in the rate of the reaction catalysed by NADP-ICDH and hence
to an additional and complementary effect on the net metabolic flux from isocitrate
to α-ketoglutarate. Other consequences of this substrate cycle are, (i) the production
of heat at the expense of Δp, which may contribute to thermoregulation in the
animal, and (ii) an increased rate of dissipation of Δp (leak).
acknowledgement: LAS is grateful to the Wellcome Trust for a fellowship. We should
like to thank Prof. R.M. Denton for discussion.
article_processing_charge: No
article_type: original
author:
- first_name: Leonid A
full_name: Sazanov, Leonid A
id: 338D39FE-F248-11E8-B48F-1D18A9856A87
last_name: Sazanov
orcid: 0000-0002-0977-7989
- first_name: Julie
full_name: Jackson, Julie
last_name: Jackson
citation:
ama: Sazanov LA, Jackson J. Proton translocating transhydrogenase and NAD- and NADP-linked
isocitrate dehydrogenases operate in a substrate cycle which contributes to fine
regulation of the tricarboxylic acid cycle activity in mitochondria. FEBS Letters.
1994;344(2-3):109-116. doi:10.1016/0014-5793(94)00370-X
apa: Sazanov, L. A., & Jackson, J. (1994). Proton translocating transhydrogenase
and NAD- and NADP-linked isocitrate dehydrogenases operate in a substrate cycle
which contributes to fine regulation of the tricarboxylic acid cycle activity
in mitochondria. FEBS Letters. Elsevier. https://doi.org/10.1016/0014-5793(94)00370-X
chicago: Sazanov, Leonid A, and Julie Jackson. “Proton Translocating Transhydrogenase
and NAD- and NADP-Linked Isocitrate Dehydrogenases Operate in a Substrate Cycle
Which Contributes to Fine Regulation of the Tricarboxylic Acid Cycle Activity
in Mitochondria.” FEBS Letters. Elsevier, 1994. https://doi.org/10.1016/0014-5793(94)00370-X.
ieee: L. A. Sazanov and J. Jackson, “Proton translocating transhydrogenase and NAD-
and NADP-linked isocitrate dehydrogenases operate in a substrate cycle which contributes
to fine regulation of the tricarboxylic acid cycle activity in mitochondria,”
FEBS Letters, vol. 344, no. 2–3. Elsevier, pp. 109–116, 1994.
ista: Sazanov LA, Jackson J. 1994. Proton translocating transhydrogenase and NAD-
and NADP-linked isocitrate dehydrogenases operate in a substrate cycle which contributes
to fine regulation of the tricarboxylic acid cycle activity in mitochondria. FEBS
Letters. 344(2–3), 109–116.
mla: Sazanov, Leonid A., and Julie Jackson. “Proton Translocating Transhydrogenase
and NAD- and NADP-Linked Isocitrate Dehydrogenases Operate in a Substrate Cycle
Which Contributes to Fine Regulation of the Tricarboxylic Acid Cycle Activity
in Mitochondria.” FEBS Letters, vol. 344, no. 2–3, Elsevier, 1994, pp.
109–16, doi:10.1016/0014-5793(94)00370-X.
short: L.A. Sazanov, J. Jackson, FEBS Letters 344 (1994) 109–116.
date_created: 2018-12-11T11:54:52Z
date_published: 1994-05-16T00:00:00Z
date_updated: 2022-06-09T13:21:50Z
day: '16'
doi: 10.1016/0014-5793(94)00370-X
extern: '1'
external_id:
pmid:
- '8187868'
intvolume: ' 344'
issue: 2-3
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://febs.onlinelibrary.wiley.com/doi/abs/10.1016/0014-5793%2894%2900370-X
month: '05'
oa: 1
oa_version: Published Version
page: 109 - 116
pmid: 1
publication: FEBS Letters
publication_identifier:
issn:
- 0014-5793
publication_status: published
publisher: Elsevier
publist_id: '5134'
quality_controlled: '1'
status: public
title: Proton translocating transhydrogenase and NAD- and NADP-linked isocitrate dehydrogenases
operate in a substrate cycle which contributes to fine regulation of the tricarboxylic
acid cycle activity in mitochondria
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 344
year: '1994'
...
---
_id: '1953'
abstract:
- lang: eng
text: The respiratory burst induced by phorbol myristate acetate in mouse macrophages
was inhibited by ultra-low doses (10-15 -10-13 M) of an opioid peptide [d-Ala2]
methionine enkephalinamide. The effect disappeared at concentrations above and
below this range. The inhibition approached 50% and was statistically significant
(P < 0.001). Increasing the time of the opioid incubation with cells brought
about a shift in the maximal effect to lower concentrations of the opioid (from
10-13 to 5 · 10-15 M) and led to a decrease in the value of the effect, fully
in accord with the previously proposed adaptation mechanism of the action of ultra-low
doses.
article_processing_charge: No
article_type: original
author:
- first_name: Alexander
full_name: Efanov, Alexander
last_name: Efanov
- first_name: Aleksei
full_name: Koshkin, Aleksei
last_name: Koshkin
- first_name: Leonid A
full_name: Sazanov, Leonid A
id: 338D39FE-F248-11E8-B48F-1D18A9856A87
last_name: Sazanov
orcid: 0000-0002-0977-7989
- first_name: O I
full_name: Borodulina, O I
last_name: Borodulina
- first_name: Sergei
full_name: Varfolomeev, Sergei
last_name: Varfolomeev
- first_name: Sergei
full_name: Zaǐtsev, Sergei
last_name: Zaǐtsev
citation:
ama: Efanov A, Koshkin A, Sazanov LA, Borodulina OI, Varfolomeev S, Zaǐtsev S. Inhibition
of the respiratory burst in mouse macrophages by ultra-low doses of an opioid
peptide is consistent with a possible adaptation mechanism. FEBS Letters.
1994;355(2):114-116. doi:10.1016/0014-5793(94)01109-5
apa: Efanov, A., Koshkin, A., Sazanov, L. A., Borodulina, O. I., Varfolomeev, S.,
& Zaǐtsev, S. (1994). Inhibition of the respiratory burst in mouse macrophages
by ultra-low doses of an opioid peptide is consistent with a possible adaptation
mechanism. FEBS Letters. Elsevier. https://doi.org/10.1016/0014-5793(94)01109-5
chicago: Efanov, Alexander, Aleksei Koshkin, Leonid A Sazanov, O I Borodulina, Sergei
Varfolomeev, and Sergei Zaǐtsev. “Inhibition of the Respiratory Burst in Mouse
Macrophages by Ultra-Low Doses of an Opioid Peptide Is Consistent with a Possible
Adaptation Mechanism.” FEBS Letters. Elsevier, 1994. https://doi.org/10.1016/0014-5793(94)01109-5.
ieee: A. Efanov, A. Koshkin, L. A. Sazanov, O. I. Borodulina, S. Varfolomeev, and
S. Zaǐtsev, “Inhibition of the respiratory burst in mouse macrophages by ultra-low
doses of an opioid peptide is consistent with a possible adaptation mechanism,”
FEBS Letters, vol. 355, no. 2. Elsevier, pp. 114–116, 1994.
ista: Efanov A, Koshkin A, Sazanov LA, Borodulina OI, Varfolomeev S, Zaǐtsev S.
1994. Inhibition of the respiratory burst in mouse macrophages by ultra-low doses
of an opioid peptide is consistent with a possible adaptation mechanism. FEBS
Letters. 355(2), 114–116.
mla: Efanov, Alexander, et al. “Inhibition of the Respiratory Burst in Mouse Macrophages
by Ultra-Low Doses of an Opioid Peptide Is Consistent with a Possible Adaptation
Mechanism.” FEBS Letters, vol. 355, no. 2, Elsevier, 1994, pp. 114–16,
doi:10.1016/0014-5793(94)01109-5.
short: A. Efanov, A. Koshkin, L.A. Sazanov, O.I. Borodulina, S. Varfolomeev, S.
Zaǐtsev, FEBS Letters 355 (1994) 114–116.
date_created: 2018-12-11T11:54:53Z
date_published: 1994-11-28T00:00:00Z
date_updated: 2022-06-09T12:58:57Z
day: '28'
doi: 10.1016/0014-5793(94)01109-5
extern: '1'
external_id:
pmid:
- '7982481'
intvolume: ' 355'
issue: '2'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://febs.onlinelibrary.wiley.com/doi/abs/10.1016/0014-5793%2894%2901109-5
month: '11'
oa: 1
oa_version: Published Version
page: 114 - 116
pmid: 1
publication: FEBS Letters
publication_identifier:
issn:
- 0014-5793
publication_status: published
publisher: Elsevier
publist_id: '5133'
quality_controlled: '1'
status: public
title: Inhibition of the respiratory burst in mouse macrophages by ultra-low doses
of an opioid peptide is consistent with a possible adaptation mechanism
type: journal_article
user_id: ea97e931-d5af-11eb-85d4-e6957dddbf17
volume: 355
year: '1994'
...