13C- and 1H-NMR studies of oxyanion and tetrahedral intermediate stabilization by the serine proteinases: optimizing inhibitor warhead specificity and potency by studying the inhibition of the serine proteinases by peptide-derived chloromethane and glyoxal inhibitors. (22nd May 2007)

• Record Type: Journal Article
• Authors: Malthouse, J.P.G.
• Is Part Of: Biochemical Society transactions. Volume 35:Number 3(2007)
• Page Start: 566
• Page End: 570
• Publication Date: 2007-05-22
• Subjects: chloromethane -- glyoxal -- inhibitor -- oxyanion -- serine proteinase -- tetrahedral intermediate; Biochemistry -- Congresses; 572
• DOI: 10.1042/BST0350566
• Languages: English
• ISSNs: 0300-5127

Abstract

Abstract : Catalysis by the serine proteinases proceeds via a tetrahedral intermediate whose oxyanion is stabilized by hydrogen-bonding in the oxyanion hole. There have been extensive 13 C-NMR studies of oxyanion and tetrahedral intermediate stabilization in trypsin, subtilisin and chymotrypsin using substrate-derived chloromethane inhibitors. One of the limitations of these inhibitors is that they irreversibly alkylate the active-site histidine residue which results in the oxyanion not being in the optimal position in the oxyanion hole. Substrate-derived glyoxal inhibitors are reversible inhibitors which, if they form tetrahedral adducts in the same way as substrates form tetrahedral intermediates, will overcome this limitation. Therefore we have synthesized 13 C-enriched substrate-derived glyoxal inhibitors which have allowed us to use 13 C-NMR and 1 H-NMR to determine how they interact with proteinases. It is hoped that these studies will help in the design of specific and highly potent warheads for serine proteinase inhibitors.