Unit

UNIT 21.22 Proteolytic Fingerprinting of Complex Biological Samples Using Combinatorial Libraries of Fluorogenic Probes

  1. Kalyani Jambunathan,
  2. Douglas S. Watson,
  3. Krishna Kodukula,
  4. Amit K. Galande

Published Online: 1 NOV 2012

DOI: 10.1002/0471140864.ps2122s70

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Jambunathan, K., Watson, D. S., Kodukula, K. and Galande, A. K. 2012. Proteolytic Fingerprinting of Complex Biological Samples Using Combinatorial Libraries of Fluorogenic Probes. Current Protocols in Protein Science. 70:21.22:21.22.1–21.22.14.

Author Information

  1. Center for Advanced Drug Research, Biosciences Division, SRI International, Harrisonburg, Virginia

Publication History

  1. Published Online: 1 NOV 2012
  2. Published Print: NOV 2012

Abstract

Proteases have garnered interest as candidate biomarkers and therapeutic targets for many human diseases. A key challenge is the identification and characterization of disease-relevant proteases in the complex milieu of biological fluids such as serum, plasma, and bronchoalveolar lavage, in which a multitude of hydrolases act in concert. This unit describes a protocol to map the global proteolytic substrate specificities of complex biological samples using a concise combinatorial library of internally quenched fluorogenic peptide probes (IQFPs). This substrate profiling approach provides a global and quantitative comparison of protease specificities between different biological samples. Such a comparative analysis can lead to the identification of disease-specific ‘fingerprints' of proteolytic activities with potential utility in diagnosis and therapy. Curr. Protoc. Protein Sci. 70:21.22.1-21.22.14. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • FRET;
  • global proteolytic specificities;
  • proteases;
  • protease substrate identification