Substrate mimetics in protease catalysis: characteristics, kinetics, and synthetic utility.
This article reviews the latest developments in protease-catalyzed peptide synthesis focusing on the use of substrate mimetics. The substrate mimetics approach takes advantage of the characteristic of this novel type of substrates to direct the enzyme to recognize an alternative site on the acyl donor, i.e. the site-specific ester leaving group, mediating the acceptance of originally poorly reactive acyl moieties. At first the kinetics and catalytic mechanism of substrate mimetics-mediated reactions are discussed on the basis of hydrolysis, peptide synthesis, protein-ligand docking, and molecular dynamics studies. By the example of the Glu-specific V8 protease and the aromatic amino acid-specific chymotrypsin both the empirical and computer-aided design of specific substrate mimetics is described. The influence of the leaving group specifically recognized by the enzyme is also considered. The benefits of these artificial substrates over common acyl donor components are illustrated by selected synthesis reactions of small peptides, peptide isosteres, non-peptidic carboxylic acid amides, and the coupling of peptide fragments at non-specific ligation sites resulting in biologically active peptide products. Finally, this review focuses on critical syntheses that uses specific-amino acid-containing peptides as the reactants of ligation. Based on these, the restrictions of the substrate mimetics approach is critically discussed and techniques to their overcoming are presented.
Bordusa
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Max-Planck Society, Research Unit Enzymology of Protein Folding, Weinbergweg 22, D-06120 Halle/Saale, Germany. bordusa@enzyme-halle.mpg.de
D000215:Acylation; D002384:Catalysis; D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism; D017076:Computer-Aided Design; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D018716:Molecular Mimicry; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D008024:Ligands; D010455:Peptides; D010450:Endopeptidases; D002918:Chymotrypsin; D014357:Trypsin