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Stimulation of Rho GDI release by ERM proteins

2000; Academic Press; Linguagem: Inglês

10.1016/s0076-6879(00)25434-7

1557-7988

Akiko Mammoto, Kazoo Takahashi, Takuya Sasaki, Yoshimi Takai,

Protein Kinase Regulation and GTPase Signaling

This chapter discusses the stimulation of Rho guanine dissociation inhibitor (GDI) released by ezrin/radixin/moesin (ERM) proteins. The Rho small guanosine 5'-triphosphates (GTPases) family is implicated in various cell functions, such as cell shape change, cell motility, and cytokinesis, through reorganization of the actin cytoskeleton. The chapter describes the procedures for the purification of recombinant full-length and truncated mutants of radixin, the isolation of the Rho A-Rho GDI complex, and the cell-free assay for radixin activity toward Rho GDI. The chapter discusses the procedures for the analysis of radixin induced activation of Rho subfamily members in intact cells. The chapter also discusses the isolation of Rho A-Rho GDI complex as well as the properties of radixin. The chapter concludes with a description of an assay for Nr-fragment-induced activation of Rho subfamily members in intact cells.