Protocol

Bioconjugation Protocols

Volume 283 of the series Methods in Molecular Biology™ pp 233-244

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

  • Thomas DurekAffiliated withMax-Planck-Institut für Molekulare Physiologie
  • , Roger S. GoodyAffiliated withMax-Planck-Institut für Molekulare Physiologie
  • , Kirill AlexandrovAffiliated withMax-Planck-Institut für Molekulare Physiologie

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Abstract

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

Key Words

Rab proteins GTPase prenylation expressed protein ligation peptides