5M6X

Crystal Structure of human RhoGAP mutated in its arginine finger (R85A) in complex with RhoA.GDP.MgF3- human

Summary for 5M6X

• Entry DOI: 10.2210/pdb5m6x/pdb
• Related: 5C4M
• Descriptor: Rho GTPase-activating protein 1, Transforming protein RhoA, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: rhogap, arginine finger, rhoa, transition state, signaling protein
• Biological source: Homo sapiens (Human); More
• Cellular location: Cytoplasm: Q07960; Cell membrane; Lipid-anchor; Cytoplasmic side: P61586
• Total number of polymer chains: 4
• Total formula weight: 98743.69

Authors

Pellegrini, E.,Bowler, M.W. (deposition date: 2016-10-26, release date: 2017-05-17, Last modification date: 2024-01-17)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.4 Å)