UNIT 14.3 Analysis of Protein Prenylation In Vitro and In Vivo Using Functionalized Phosphoisoprenoids

  1. Uyen T.T. Nguyen1,
  2. Yaowen Wu2,
  3. Andrew Goodall3,
  4. Kirill Alexandrov3

Published Online: 1 NOV 2010

DOI: 10.1002/0471140864.ps1403s62

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Nguyen, U. T., Wu, Y., Goodall, A. and Alexandrov, K. 2010. Analysis of Protein Prenylation In Vitro and In Vivo Using Functionalized Phosphoisoprenoids. Current Protocols in Protein Science. 62:14.3:14.3.1–14.3.15.

Author Information

  1. 1

    Max Planck Institute, Dortmund, Germany

  2. 2

    Department of Physical Biochemistry, Max-Planck Institute of Molecular Physiology, Dortmund, Germany

  3. 3

    Department of Cell and Molecular Biology, Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Australia

Publication History

  1. Published Online: 1 NOV 2010
  2. Published Print: NOV 2010


Post-translational modifications (PTMs) expand the number of protein isoforms in eukaryotic proteome by orders of magnitude. Protein modification with isoprenoid lipids is a common PTM affecting hundreds of proteins controlling the transport of information and materials into, through, and out of the eukaryotic cell. In this modification, a soluble phosphoisoprenoid such as farnesyl (C15) or geranylgeranyl (C20) pyrophosphate moiety is recruited by one of three protein prenyltransferases to covalently modify a C-terminal cysteine(s) in a target protein. The three mammalian prenyltransferases are farnesyltransferase (FTase), geranylgeranyltransferase type I (GGTase I), and Rab geranylgeranyl transferase (also termed geranylgeranyltransferase type II - GGTase II). In this unit, synthetic isoprenoids conjugated to either a fluorophore or biotin group are used to assay the activity of protein prenyltransferases in vitro or to affinity tag prenylatable proteins in cell lysates. These protocols and their modifications can be used to study the mechanisms of protein prenylation, identify prenylation targets, and characterize inhibitors of protein prenyltransferases in vitro and in vivo. Curr. Protoc. Protein Sci. 62:14.3.1-14.3.15. © 2010 by John Wiley & Sons, Inc.


  • protein prenylation;
  • isoprenoids;
  • protein prenyltransferases