4PHH

Crystal structure of Ypt7 covalently modified with GNP

Summary for 4PHH

• Entry DOI: 10.2210/pdb4phh/pdb
• Related: 4PHF 4PHG
• Descriptor: GTP-binding protein YPT7, 5'-O-[(R)-hydroxy{[(S)-hydroxy(phosphonoamino)phosphoryl]oxy}phosphoryl]-N-[3-(propanoylamino)propyl]guanosine, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: ypt7, acryl-nucleotides, agnp, covalent, gnp, gppnhp, endocytosis, exocytosis
• Biological source: Saccharomyces cerevisiae (Baker's yeast)
• Total number of polymer chains: 4
• Total formula weight: 84064.88

Authors

Wiegandt, D.,Vieweg, S.,Hofmann, F.,Koch, D.,Wu, Y.,Itzen, A.,Mueller, M.P.,Goody, R.S. (deposition date: 2014-05-06, release date: 2014-05-28, Last modification date: 2024-10-23)

Primary citation

Wiegandt, D.,Vieweg, S.,Hofmann, F.,Koch, D.,Li, F.,Wu, Y.W.,Itzen, A.,Muller, M.P.,Goody, R.S.
Locking GTPases covalently in their functional states.
Nat Commun, 6:7773-7773, 2015

PubMed 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.

PubMed: 26178622
DOI: 10.1038/ncomms8773

Experimental method

X-RAY DIFFRACTION (2.35 Å)