title: Mapping the site of interaction between annexin VI and the p120GAP C2 domain.
abstract: Annexin VI is a Ca(2+)-dependent membrane and phospholipid binding protein. It mediates a protein-protein interaction with the Ras p21 regulatory protein p120GAP. In this study we have mapped the binding site of GAP within the annexin VI protein. Using Far Western overlay binding assays and cell lysate competition studies we have mapped the site of interaction to the inter-lobe linker region; amino acids 325-363. Finally, using a GST fusion protein corresponding to this linker region we have demonstrated that cellular loading of the fusion protein into Rat-1 fibroblasts by electroporation blocks the interaction and co-immunoprecipitation of annexin VI and GAP.
authors.affiliation: School of Biomedical Sciences, University of Leeds, Yorkshire, UK.
authors.forename: A
authors.initials: A
authors.lastname: Chow
authors.forename: D
authors.initials: D
authors.lastname: Gawler
mesh_terms: D000818:Animals; D017317:Annexin A6 / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D001667:Binding, Competitive; D002460:Cell Line; D018274:Electroporation; D004926:Escherichia coli; D006801:Humans; D010446:Peptide Fragments / Q000494:pharmacology; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012697:Serine Endopeptidases; D020729:p120 GTPase Activating Protein / Q000737:chemistry* / Q000235:genetics
chemical_list: D017317:Annexin A6; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D020729:p120 GTPase Activating Protein; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase