Assay and properties of the GIT1/p95-APP1 ARFGAP.
GIT1/p95-APP1 is an adaptor protein with an aminoterminal ARFGAP domain involved in the regulation of ARF6 function. GIT1/p95-APP1 forms stable complexes with a number of proteins including downstream effectors and exchanging factors for members of the Rho family of small GTPases. This protein can also interact with other adaptor proteins implicated in the regulation of cell adhesion and synapse formation. The stability of the endogenous and reconstituted complexes after cell lysis allows the biochemical identification and characterization of the GIT1 complexes that can be isolated from different cell types. This article presents methods for the identification of the endogenous and reconstituted GIT1 complexes that can be utilized for the biochemical and functional characterization of the complexes from different tissue and cell types.
de Curtis
Ivan
I
San Rafaele Scientific Institute, Department of Molecular Biology and Functional Genomics, Milano, Italy.
Paris
Simona
S
D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D018797:Cell Cycle Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D002846:Chromatography, Affinity / Q000379:methods; D020690:GTPase-Activating Proteins / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D064067:Rho Guanine Nucleotide Exchange Factors; D020744:rac GTP-Binding Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D000089982:ADP-Ribosylation Factor 6; D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; C485647:GIT1 protein, human; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D010750:Phosphoproteins; D064067:Rho Guanine Nucleotide Exchange Factors; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human; D020744:rac GTP-Binding Proteins