Unit

UNIT 3.11 Quantitative Analysis of Surface Expression of Membrane Proteins Using Cold-Adapted Proteases

  1. Faraz Ahmad1,
  2. Kai Kaila1,
  3. Peter Blaesse1,2

Published Online: 24 SEP 2013

DOI: 10.1002/0471140864.ps0311s73

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Ahmad, F., Kaila, K. and Blaesse, P. 2013. Quantitative Analysis of Surface Expression of Membrane Proteins Using Cold-Adapted Proteases. Current Protocols in Protein Science. 73:3.11:3.11.1–3.11.12.

Author Information

  1. 1

    Department of Biosciences and Neuroscience Center, University of Helsinki, Helsinki, Finland

  2. 2

    Institute of Physiology I, Westfälische Wilhelms-University Münster, Münster, Germany

Publication History

  1. Published Online: 24 SEP 2013

Abstract

This unit presents an improved method for quantitative analysis of surface expression of membrane proteins utilizing a cold-adapted trypsin. Preservation of the proteolytic activity of the enzyme at 0° to 4°C allows cleavage of surface-expressed membrane proteins at temperatures at which trafficking of the mammalian plasmalemmal proteins is blocked. This provides an important advantage over established trypsin-cleavage protocols since it can be applied to membrane proteins with a fast turnover rate of the membrane pool and a fast recycling rate. Compared to surface biotinylation, the method is less time consuming. Curr. Protoc. Protein Sci. 73:3.11.1-3.11.12. © 2013 by JohnWiley & Sons, Inc.

Keywords:

  • membrane protein;
  • surface expression;
  • trafficking;
  • trypsin;
  • biotinylation;
  • BS3;
  • surface analysis