Scanning the prime-site substrate specificity of proteolytic enzymes

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Subtitle: A novel assay based on ligand-enhanced lanthanide ion fluorescence
Author list: Barrios AM, Craik CS
Publisher: Elsevier
Publication year: 2002
Volume number: 12
Issue number: 24
Start page: 3619
End page: 3623
Number of pages: 5
ISSN: 0960-894X
Languages: English-Great Britain (EN-GB)


Abstract

A novel method for assaying the substrate specificity of proteolytic enzymes has been developed utilizing ligand-enhanced lanthanide ion fluorescence. This approach was used to develop peptide libraries to probe substrate specificity in the prime sites of proteolytic enzymes. A positional scanning synthetic combinatorial library of fluorogenic peptides was synthesized and used to determine the extended prime site specificity of bovine alpha-chymotrypsin. The enzyme showed a preference for Lys and Arg in the P1' position, rather broad specificity in the P2' position, and a slight Arg specificity in the P3' position. The specificity profile of bovine alpha-chymotrypsin agrees well with previously reported data, and the substrate library reported herein should provide valuable information about the prime site substrate specificities of other proteolytic enzymes as well. Furthermore, the continuous fluorogenic assay described may prove useful in analyzing the activity of other hydrolytic enzymes. (C) 2002 Elsevier Science Ltd. All rights reserved.


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Last updated on 2019-23-08 at 11:15