Unit

UNIT 14.3 Analysis of Protein Prenylation and Carboxyl-Methylation

  1. Anthony I. Magee

Published Online: 1 MAY 2001

DOI: 10.1002/0471140864.ps1403s05

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Magee, A. I. 2001. Analysis of Protein Prenylation and Carboxyl-Methylation. Current Protocols in Protein Science. 5:14.3:14.3.1–14.3.11.

Author Information

  1. National Institute for Medical Research, London, United Kingdom

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: SEP 1996

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Abstract

This unit describes methods for analysis of prenylation and the carboxyl-methylation that often accompanies it. The two prenoid groups that have been found attached to proteins--farnesyl (C15) and geranylgeranyl (C20)--are both derived from intermediates in the isoprenoid biosynthetic pathway that utilizes mevalonic acid. In the protocols described here, radiolabeled mevalonate is used to label these intermediates in either intact cells or in vitro; the labeled intermediates then become incorporated into proteins. Alternatively, the preformed radioactive prenyl diphosphates can be used for in vitro translations, as described here. Carboxyl-methylation of C-terminal prenylated cysteine residues often occurs subsequent to prenylation. Methods are given for radiolabeling of the methyl group with [3H-methyl]methionine, that is converted intracellularly into S-adenosylmethionine, and for radiolabeling with preformed S-adenosyl[3H]methionine.