Research ExplorerPublicationsLocking GTPases covalently in their functional states

Locking GTPases covalently in their functional states

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Locking GTPases covalently in their functional states. / Wiegandt, David; Vieweg, Sophie; Hofmann, Frank; Koch, Daniel; Li, Fu; Wu, Yao-Wen; Itzen, Aymelt; Goody, Roger S.

In: NAT COMMUN, Vol. 6, 16.07.2015, p. 7773.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Wiegandt, D, Vieweg, S, Hofmann, F, Koch, D, Li, F, Wu, Y-W, Itzen, A & Goody, RS 2015, 'Locking GTPases covalently in their functional states', NAT COMMUN, vol. 6, pp. 7773. https://doi.org/10.1038/ncomms8773

APA

Wiegandt, D., Vieweg, S., Hofmann, F., Koch, D., Li, F., Wu, Y-W., Itzen, A., & Goody, R. S. (2015). Locking GTPases covalently in their functional states. NAT COMMUN, 6, 7773. https://doi.org/10.1038/ncomms8773

Vancouver

Wiegandt D, Vieweg S, Hofmann F, Koch D, Li F, Wu Y-W et al. Locking GTPases covalently in their functional states. NAT COMMUN. 2015 Jul 16;6:7773. https://doi.org/10.1038/ncomms8773

Bibtex

@article{5a05d8456f26483d9b532826b3556f79,
title = "Locking GTPases covalently in their functional states",
abstract = "GTPases act as key regulators of many cellular processes by switching between active (GTP-bound) and inactive (GDP-bound) states. In many cases, understanding their mode of action has been aided by artificially stabilizing one of these states either by designing mutant proteins or by complexation with non-hydrolysable GTP analogues. Because of inherent disadvantages in these approaches, we have developed acryl-bearing GTP and GDP derivatives that can be covalently linked with strategically placed cysteines within the GTPase of interest. Binding studies with GTPase-interacting proteins and X-ray crystallography analysis demonstrate that the molecular properties of the covalent GTPase-acryl-nucleotide adducts are a faithful reflection of those of the corresponding native states and are advantageously permanently locked in a defined nucleotide (that is active or inactive) state. In a first application, in vivo experiments using covalently locked Rab5 variants provide new insights into the mechanism of correct intracellular localization of Rab proteins. ",
keywords = "Binding Sites, Crystallography, X-Ray, Escherichia coli, Escherichia coli Proteins, Fungal Proteins, GTP Phosphohydrolases, Guanosine Diphosphate, Guanosine Triphosphate, Protein Binding, rab GTP-Binding Proteins, Journal Article, Research Support, Non-U.S. Gov't",
author = "David Wiegandt and Sophie Vieweg and Frank Hofmann and Daniel Koch and Fu Li and Yao-Wen Wu and Aymelt Itzen and Goody, {Roger S}",
year = "2015",
month = jul,
day = "16",
doi = "10.1038/ncomms8773",
language = "English",
volume = "6",
pages = "7773",
journal = "NAT COMMUN",
issn = "2041-1723",
publisher = "NATURE PUBLISHING GROUP",

}

RIS

TY - JOUR

T1 - Locking GTPases covalently in their functional states

AU - Wiegandt, David

AU - Vieweg, Sophie

AU - Hofmann, Frank

AU - Koch, Daniel

AU - Li, Fu

AU - Wu, Yao-Wen

AU - Itzen, Aymelt

AU - Goody, Roger S

PY - 2015/7/16

Y1 - 2015/7/16

N2 - GTPases act as key regulators of many cellular processes by switching between active (GTP-bound) and inactive (GDP-bound) states. In many cases, understanding their mode of action has been aided by artificially stabilizing one of these states either by designing mutant proteins or by complexation with non-hydrolysable GTP analogues. Because of inherent disadvantages in these approaches, we have developed acryl-bearing GTP and GDP derivatives that can be covalently linked with strategically placed cysteines within the GTPase of interest. Binding studies with GTPase-interacting proteins and X-ray crystallography analysis demonstrate that the molecular properties of the covalent GTPase-acryl-nucleotide adducts are a faithful reflection of those of the corresponding native states and are advantageously permanently locked in a defined nucleotide (that is active or inactive) state. In a first application, in vivo experiments using covalently locked Rab5 variants provide new insights into the mechanism of correct intracellular localization of Rab proteins.

AB - GTPases act as key regulators of many cellular processes by switching between active (GTP-bound) and inactive (GDP-bound) states. In many cases, understanding their mode of action has been aided by artificially stabilizing one of these states either by designing mutant proteins or by complexation with non-hydrolysable GTP analogues. Because of inherent disadvantages in these approaches, we have developed acryl-bearing GTP and GDP derivatives that can be covalently linked with strategically placed cysteines within the GTPase of interest. Binding studies with GTPase-interacting proteins and X-ray crystallography analysis demonstrate that the molecular properties of the covalent GTPase-acryl-nucleotide adducts are a faithful reflection of those of the corresponding native states and are advantageously permanently locked in a defined nucleotide (that is active or inactive) state. In a first application, in vivo experiments using covalently locked Rab5 variants provide new insights into the mechanism of correct intracellular localization of Rab proteins.

KW - Binding Sites

KW - Crystallography, X-Ray

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Fungal Proteins

KW - GTP Phosphohydrolases

KW - Guanosine Diphosphate

KW - Guanosine Triphosphate

KW - Protein Binding

KW - rab GTP-Binding Proteins

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1038/ncomms8773

DO - 10.1038/ncomms8773

M3 - SCORING: Journal article

C2 - 26178622

VL - 6

SP - 7773

JO - NAT COMMUN

JF - NAT COMMUN

SN - 2041-1723

ER -