Unit

UNIT 26.5 TimeSTAMP Tagging of Newly Synthesized Proteins

  1. Michael Z. Lin1,
  2. Roger Y. Tsien2

Published Online: 1 FEB 2010

DOI: 10.1002/0471140864.ps2605s59

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Lin, M. Z. and Tsien, R. Y. 2010. TimeSTAMP Tagging of Newly Synthesized Proteins. Current Protocols in Protein Science. 59:26.5:26.5.1–26.5.11.

Author Information

  1. 1

    Department of Pediatrics and Programs in Gene Therapy and Molecular Imaging, Stanford University, Stanford, California

  2. 2

    Department of Pharmacology and Howard Hughes Medical Institute, University of California San Diego, La Jolla, California

Publication History

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

Abstract

The ability to quantify or visualize newly synthesized proteins has important uses in cell biology. For example, a researcher may wish to quantify basal or inducible rates of translation of a specific gene of interest, or detect subcellular locations of newly synthesized copies of a protein in order to study the role of new protein synthesis in the growth of specialized cellular structures. In this unit, the TimeSTAMP method for labeling of newly synthesized copies of a protein of interest is described. In the TimeSTAMP method, the experimenter expresses a protein of interest as a fusion with a cis-acting protease and an epitope tag, both of which are removed by default protease activity. Addition of a specific protease inhibitor then allows preservation of the tag on subsequently synthesized proteins. Finally, the tag is detected by immunological methods. Curr. Protoc. Protein Sci. 59:26.5.1-26.5.11. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • protein synthesis;
  • protease;
  • conditional labeling