In Vitro Semisynthesis and Applications of C-Terminally Modified Rab Proteins

Expressed protein ligation is a powerful tool for the generation of natively folded proteins composed of recombinantly generated and chemically synthesized polypeptides. Using this approach, we developed protocols for the production of prenylated and/or otherwise-labeled Rab GTPase. The protocols are generally applicable to most small GTPases that can be supplied with a variety of new chemical functionalities. We used semisynthetic fluorescently labeled Rab7 GTPase as a molecular probe to study protein-protein interactions with components of the prenylation machinery