4phg

Publication Abstract from PubMed

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.

Locking GTPases covalently in their functional states.,Wiegandt D, Vieweg S, Hofmann F, Koch D, Li F, Wu YW, Itzen A, Muller MP, Goody RS Nat Commun. 2015 Jul 16;6:7773. doi: 10.1038/ncomms8773. PMID:26178622^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.