Unit

UNIT 20.11 Quantitative Determination of Protein Stability and Ligand Binding by Pulse Proteolysis

  1. Chiwook Park1,
  2. Susan Marqusee2

Published Online: 1 DEC 2006

DOI: 10.1002/0471140864.ps2011s46

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Park, C. and Marqusee, S. 2006. Quantitative Determination of Protein Stability and Ligand Binding by Pulse Proteolysis. Current Protocols in Protein Science. 46:20.11:20.11.1–20.11.14.

Author Information

  1. 1

    Purdue University, West Lafayette, Indiana

  2. 2

    University of California, Berkeley, California

Publication History

  1. Published Online: 1 DEC 2006
  2. Published Print: NOV 2006

Abstract

Pulse proteolysis exploits the difference in proteolytic susceptibility between folded and unfolded proteins for facile but quantitative determination of protein stability. The method requires only common biochemistry and molecular biology lab equipment. Pulse proteolysis also can be used to determine the affinity of a ligand to its protein target by monitoring the change in protein stability upon ligand binding. The Basic Protocol describes the detailed procedure for determining protein stability using pulse proteolysis. For pulse proteolysis to be used for determining a protein's stability, the protein should not be digested significantly by pulse proteolysis when it is in the folded conformation. The Support Protocol describes a procedure for determining whether a protein satisfies this requirement. The principles of protein stability determination using denaturant and pulse proteolysis are also discussed.

Keywords:

  • protein stability;
  • proteolysis;
  • ligand binding