Affiliation:
1. Biomolecular Sciences Group, School of Pharmacy, The Queen's University of Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, U.K.
Abstract
We report on the first synthesis, kinetic evaluation and application of novel substrate-derived inhibitors against the Staphylococcus aureus cysteine protease-transpeptidase, sortase (staphylococcal surface protein sorting A, SrtA). The peptidyl-diazomethane and peptidyl-chloromethane analogues, Cbz (benzyloxycarbonyl)-Leu-Pro-Ala-Thr-CHN2 (I) and Cbz-Leu-Pro-Ala-Thr-CH2Cl (II) respectively were found to act as time-dependent irreversible inhibitors of recombinant sortase (SrtAΔN). The peptidyl-chloromethane analogue (II) was the most powerful with an inhibitor specificity constant (ki/Ki) of 5.3×104M-1·min-1, approx. 2-fold greater than that determined for the peptidyl-diazomethane (I). Additionally, using Western-blot analysis, we have been able to demonstrate that a biotinylated version of the peptidyl-diazomethane analogue, biotin-Ahx (aminohexanoyl)-Leu-Pro-Ala-Thr-CHN2 (III), can be used as an affinity label to detect the presence of wild-type SrtA in crude cell lysates prepared from S. aureus.
Subject
Cell Biology,Molecular Biology,Biochemistry
Cited by
74 articles.
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