PDBsum entry 5irc

Protein binding

PDB id: 5irc

Contents

Protein chains

201 a.a.

170 a.a.

Ligands

GDP ×2

MGF ×2

Metals

_CL ×3

_MG ×2

Waters ×1019

PDB id:

5irc

Name:

Protein binding

Title:

P190a gap domain complex with rhoa

Structure:

Rho gtpase-activating protein 35. Chain: a, b. Synonym: gap-associated protein p190,glucocorticoid receptor DNA- binding factor 1. Engineered: yes. Transforming protein rhoa. Chain: f, d. Synonym: rho cdna clone 12,h12. Engineered: yes

Source:

Rattus norvegicus. Rat. Organism_taxid: 10116. Gene: arhgap35, grlf1. Expressed in: escherichia coli. Expression_system_taxid: 469008. Homo sapiens. Human. Organism_taxid: 9606.

Resolution:

1.72Å R-factor: 0.171 R-free: 0.213

Authors:

U.Derewenda,Z.Derewenda

Key ref:

E.Amin et al. (2016). Deciphering the Molecular and Functional Basis of RHOGAP Family Proteins: A SYSTEMATIC APPROACH TOWARD SELECTIVE INACTIVATION OF RHO FAMILY PROTEINS. J Biol Chem, 291, 20353-20371. PubMed id: 27481945 DOI: 10.1074/jbc.M116.736967

Date:

12-Mar-16 Release date: 17-Aug-16

Protein chains

P81128  (RHG35_RAT) -  Rho GTPase-activating protein 35 from Rattus norvegicus

Seq: 1499 a.a.

Struc: 201 a.a.*

Protein chains

P61586  (RHOA_HUMAN) -  Transforming protein RhoA from Homo sapiens

Seq: 193 a.a.

Struc: 170 a.a.*

* PDB and UniProt seqs differ at 6 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains F, D: E.C.3.6.5.2 - small monomeric GTPase.
• Reaction: GTP + H2O = GDP + phosphate + H⁺

GTP + H2O = GDP (Bound ligand (Het Group name = GDP) corresponds exactly) + phosphate + H(+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1074/jbc.M116.736967

J Biol Chem 291:20353-20371 (2016)

PubMed id: 27481945

Deciphering the Molecular and Functional Basis of RHOGAP Family Proteins: A SYSTEMATIC APPROACH TOWARD SELECTIVE INACTIVATION OF RHO FAMILY PROTEINS.

E.Amin, M.Jaiswal, U.Derewenda, K.Reis, K.Nouri, K.T.Koessmeier, P.Aspenström, A.V.Somlyo, R.Dvorsky, M.R.Ahmadian.

ABSTRACT

RHO GTPase-activating proteins (RHOGAPs) are one of the major classes of regulators of the RHO-related protein family that are crucial in many cellular processes, motility, contractility, growth, differentiation, and development. Using database searches, we extracted 66 distinct human RHOGAPs, from which 57 have a common catalytic domain capable of terminating RHO protein signaling by stimulating the slow intrinsic GTP hydrolysis (GTPase) reaction. The specificity of the majority of the members of RHOGAP family is largely uncharacterized. Here, we comprehensively investigated the sequence-structure-function relationship between RHOGAPs and RHO proteins by combining our in vitro data with in silico data. The activity of 14 representatives of the RHOGAP family toward 12 RHO family proteins was determined in real time. We identified and structurally verified hot spots in the interface between RHOGAPs and RHO proteins as critical determinants for binding and catalysis. We have found that the RHOGAP domain itself is nonselective and in some cases rather inefficient under cell-free conditions. Thus, we propose that other domains of RHOGAPs confer substrate specificity and fine-tune their catalytic efficiency in cells.