Guanine nucleotide exchange catalyzed by dbl oncogene product

摘要: Publisher Summary This chapter describes methods for the expression of Dbl and homology (DH) domain in baculovirus systems discusses vitro assays to demonstrate that its DH contain guanine nucleotide exchange factors (GEFs) activity Cdc42Hs Rho GTP-binding proteins. GEFs activate Ras family members proteins by accelerating their rate GDP release and, therefore, facilitating GTP binding cells. Unlike case Ras–GEFs which vivo activities can be examined extent conversion Ras-bound immunoprecipitation thin-layer chromatography separation bound nucleotides, GEF Rho-type GTPases are difficult determine this manner because fast hydrolysis rates presence abundant Rho–GDP-dissociation inhibitor (GDI), renders most cytosolic blocks action. Thus, reconstituted assay is indispensable detecting quantitating potential molecules.