Unit

UNIT 21.9 Use of GFP as a Reporter for the Analysis of Sequence-Specific Proteases

  1. Gautam Sarath1,
  2. Steven D. Schwartzbach2

Published Online: 1 FEB 2002

DOI: 10.1002/0471140864.ps2109s26

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Sarath, G. and Schwartzbach, S. D. 2002. Use of GFP as a Reporter for the Analysis of Sequence-Specific Proteases. Current Protocols in Protein Science. 26:21.9:21.9.1–21.9.10.

Author Information

  1. 1

    University of Nebraska, Lincoln, Nebraska

  2. 2

    University of Memphis, Memphis, Tennessee

Publication History

  1. Published Online: 1 FEB 2002
  2. Published Print: DEC 2001

Abstract

This unit describes a rapid fluorescent assay for sequence-specific proteases. A recombinant His-tagged substrate-GFP fusion protein containing the sequence-specific protease-recognition sequence is used as substrate. Batch metal-chelate chromatography separates uncleaved substrate-GFP fusion protein from GFP released by proteolysis and proteolytic activity is determined by measuring the fluorescence of GFP remaining in solution.