title: The structure of the GTPase-activating domain from p50rhoGAP.
abstract: Members of the Rho family of small G proteins transduce signals from plasma-membrane receptors and control cell adhesion, motility and shape by actin cytoskeleton formation. They also activate other kinase cascades. Like all other GTPases, Rho proteins act as molecular switches, with an active GTP-bound form and an inactive GDP-bound form. The active conformation is promoted by guanine-nucleotide exchange factors, and the inactive state by GTPase-activating proteins (GAPs) which stimulate the intrinsic GTPase activity of small G proteins. Rho-specific GAP domains are found in a wide variety of large, multi-functional proteins. Here we report the crystal structure of an active 242-residue C-terminal fragment of human p50rhoGAP. The structure is an unusual arrangement of nine alpha-helices, the core of which includes a four-helix bundle. Residues conserved across the rhoGAP family are largely confined to one face of this bundle, which may be an interaction site for target G proteins. In particular, we propose that Arg 85 and Asn 194 are involved in binding G proteins and enhancing GTPase activity.
authors.affiliation: National Institute for Medical Research, London, UK.
authors.forename: T
authors.initials: T
authors.lastname: Barrett
authors.forename: B
authors.initials: B
authors.lastname: Xiao
authors.forename: E J
authors.initials: EJ
authors.lastname: Dodson
authors.forename: G
authors.initials: G
authors.lastname: Dodson
authors.forename: S B
authors.initials: SB
authors.lastname: Ludbrook
authors.forename: K
authors.initials: K
authors.lastname: Nurmahomed
authors.forename: S J
authors.initials: SJ
authors.lastname: Gamblin
authors.forename: A
authors.initials: A
authors.lastname: Musacchio
authors.forename: S J
authors.initials: SJ
authors.lastname: Smerdon
authors.forename: J F
authors.initials: JF
authors.lastname: Eccleston
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000737:chemistry*; D020690:GTPase-Activating Proteins; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D020690:GTPase-Activating Proteins; C085379:rho GTPase-activating protein; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins