Abstract
This chapter describes the process of activation of Rac1 by human T-lymphoma invasion and metastasis 1 (Tiam1). The small GTPase Rac1 becomes activated via catalyzed displacement of bound GDP by a guanine nucleotide exchange factor (GEF). There are various Tiaml homologs with very similar domain organization. All these proteins share two pleckstrin homology (PH) domains that putatively bind to lipids, a PDZ domain that typically interacts with protein partners, and a Dbl homology (DH) domain that encodes the GEF active site. The chapter describes the characterization of recombinant human Tiaml expressed in Sf9 cells and assayed in the presence of one of these amphiphilic molecules, ascorbyl stearate. The chapter also discusses the expression and purification of recombinant proteins and various assays of nucleotide exchange. The chapter concludes with a discussion of the characterization of Tiaml nucleotide exchange activity.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
