Gel electrophoresis of proteolytic enzymes Review Article

This paper reviews the major procedures devised to efficiently detect and analyze proteolytic enzymes and their inhibitors after gel electrophoresis. After some considerations on the extraction of these proteins from biological samples and on the choice of appropriate substrates for the detection of protease activities, the major analytical schemes developed to detect electrophoretically separated proteases are described. Given the well-known importance of protein protease inhibitors in the regulation of endogenous proteolytic enzymes and in host–pathogen and host–predator interactions, the detection of these proteins in crude extracts by the combined use of gel electrophoresis and specific proteases is then considered. The use of gel electrophoresis in the study of protease/protease inhibitor complexes, useful in characterizing protease mechanistic classes and in monitoring the activity of recombinant proteinaceous inhibitors designed to regulate proteases in various biological systems, is finally considered.