5M70

Crystal Structure of human RhoGAP mutated in its arginin finger (R85A) in complex with RhoA.GDP.AlF4- human

5M70 の概要

• エントリーDOI: 10.2210/pdb5m70/pdb
• 分子名称: Rho GTPase-activating protein 1, Transforming protein RhoA, MAGNESIUM ION, ... (6 entities in total)
• 機能のキーワード: rhogap, arginine finger, rhoa, transition state, signaling protein
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Cytoplasm: Q07960; Cell membrane; Lipid-anchor; Cytoplasmic side: P61586
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 98787.04

構造登録者

Pellegrini, E.,Bowler, M.W. (登録日: 2016-10-26, 公開日: 2017-05-17, 最終更新日: 2024-01-17)

主引用文献

Jin, Y.,Molt, R.W.,Pellegrini, E.,Cliff, M.J.,Bowler, M.W.,Richards, N.G.J.,Blackburn, G.M.,Waltho, J.P.
Assessing the Influence of Mutation on GTPase Transition States by Using X-ray Crystallography, (19) F NMR, and DFT Approaches.
Angew. Chem. Int. Ed. Engl., 56:9732-9735, 2017

PubMed Abstract: We report X-ray crystallographic and F NMR studies of the G-protein RhoA complexed with MgF , GDP, and RhoGAP, which has the mutation Arg85'Ala. When combined with DFT calculations, these data permit the identification of changes in transition state (TS) properties. The X-ray data show how Tyr34 maintains solvent exclusion and the core H-bond network in the active site by relocating to replace the missing Arg85' sidechain. The F NMR data show deshielding effects that indicate the main function of Arg85' is electronic polarization of the transferring phosphoryl group, primarily mediated by H-bonding to O and thence to P . DFT calculations identify electron-density redistribution and pinpoint why the TS for guanosine 5'-triphosphate (GTP) hydrolysis is higher in energy when RhoA is complexed with RhoGAP relative to wild-type (WT) RhoGAP. This study demonstrates that F NMR measurements, in combination with X-ray crystallography and DFT calculations, can reliably dissect the response of small GTPases to site-specific modifications.

PubMed: 28498638
DOI: 10.1002/anie.201703074

実験手法

X-RAY DIFFRACTION (2.2 Å)