Chapter category: Agricultural Biotechnology
Appendix I. Substrates and Inhibitors Useful in Protease Characterization
Recombinant Protease Inhibitors in Plants
Edited by: Dominique MichaudISBN: 1-58706-007-8
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Chapter authors:
France Brunelle and Dominique Michaud
substrate a chromogenic or fluorogenic nature useful in subsequent quantitation of hydrolytic activity. Based on the position of the substituent(s), synthetic substrates may be classified in two groups: endoprotease (or proteinase) substrates, in which both the amino and carboxy termini are blocked and thereby not available to exoproteases; and exoprotease substrates, in which only one extremity is blocked, the carboxyterminal end for aminopeptidases or the amino terminal end for carboxypeptidases. The most common substituents added to block NH2 extremities are the benzoyl (Bz), benzo- yloxycarbonyl (Z), acetyl (Ac) and succinyl (Suc) functional groups, while those added to COOH extremities, which give in most instances the substrate its chromogenic or fluorogenic potential, are 4nitroanilide, a chromogenic substituent, 7amido4methyl- coumarin, a fluorogenic substituent, and 2naphtylamide, a substituent exhibiting either chromogenic or fluorogenic potential, depending on the detection procedure used to visualize the activity.2 The chromogenic detection of proteases primarily depends on the difference in molar absorptivity between the substrate and its hydrolytic products while fluorogenic substrates, generally more sensitive, contain a substituent group exhibiting measurable fluorescence when separated from the intact substrate.2,3
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Appendix III. Tertiary Structures of Proteases and Protease Inhibitors Available in the Brookhaven and National Laboratory Protein Data Bank
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The GenBank Database was searched with the Wisconsin Package, Version 9.1 (Madison Genetics Computer Group, Madison WI), using the following keywords: protease inhibitor, proteinase inhibitor,...
Appendix I. Substrates and Inhibitors Useful in Protease Characterization
France Brunelle and Dominique Michaud
substrate a chromogenic or fluorogenic nature useful in subsequent quantitation of hydrolytic activity. Based on the position of the substituent(s), synthetic substrates may be classified in t...
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