title: Cloning and characterization of a rhoGAP homolog from Dictyostelium discoideum.
abstract: Small GTPases interact with a variety of proteins that affect nucleotide binding and cleavage. GTPase activating proteins (GAPs) are one class of these proteins that act by accelerating the intrinsic GTPase rate resulting in the formation of the biologically inactive GDP-bound form of the GTPase. For the Rho subfamily of GTPases, there is a growing number of proteins with rhoGAP activity that are identifiable by a homologous region of about 150 amino acids. We have exploited this homology using the polymerase chain reaction to clone the first rhoGAP homolog, called DdRacGAP, from the slime mold Dictyostelium discoideum. The GAP domain of DdRacGAP (amino acids 1-212), when expressed and purified from Escherichia coli, is active on both Dictyostelium and human Rho family GTPases but not human Ras. The full-length protein is 1356 amino acids in length and has several interesting homologies in addition to the GAP domain, including an SH3 domain, a dbl homology domain, and a pleckstrin homology domain.
authors.affiliation: National Institute for Medical Research, Department of Physical Biochemistry, The Ridgeway, Mill Hill, NW7 1AA, London, United Kingdom.
authors.forename: S B
authors.initials: SB
authors.lastname: Ludbrook
authors.forename: J F
authors.initials: JF
authors.lastname: Eccleston
authors.forename: M
authors.initials: M
authors.lastname: Strom
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004023:Dictyostelium; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D020690:GTPase-Activating Proteins; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016415:Sequence Alignment; D013379:Substrate Specificity; D018909:src Homology Domains
chemical_list: D020690:GTPase-Activating Proteins; C085379:rho GTPase-activating protein; D019204:GTP-Binding Proteins