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

Thomas Durek,Roger S. Goody,Kirill Alexandrov-2004-06-15-Humana Press eBooks
6

TL;DRAbstract

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

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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

Keywords

RabSemisynthesisGTPasePrenylationChemistryCell biologyBiochemistryIn vitro

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