In vitro semisynthesis and applications of C-terminally modified rab proteins

Methods Mol Biol. 2004;283:233-44. doi: 10.1385/1-59259-813-7:233.

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

MeSH terms

  • Electrophoresis, Polyacrylamide Gel
  • In Vitro Techniques
  • Protein Processing, Post-Translational*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Staining and Labeling
  • rab GTP-Binding Proteins / chemical synthesis*
  • rab GTP-Binding Proteins / chemistry

Substances

  • rab7 protein
  • rab GTP-Binding Proteins