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. 1992 May 1;283(Pt 3):871–876. doi: 10.1042/bj2830871

Peptidyldiazomethanes. A novel mechanism of interaction with prolyl endopeptidase.

S R Stone 1, D Rennex 1, P Wikstrom 1, E Shaw 1, J Hofsteenge 1
PMCID: PMC1130967  PMID: 1590775

Abstract

Peptidyldiazomethanes with proline in the P1 position were found to be competitive slow-binding inhibitors of prolyl endopeptidase. Progress-curve experiments monitoring the increase in the degree of inhibition with time indicated that the kinetic mechanism involved an initial complex that isomerized to form a tighter complex. Reversibility of the inhibited complex was demonstrated by monitoring the regain of enzyme activity after removal of free inhibitor and dilution into an assay containing competing substrate. The kinetics of the reversal of inhibition indicated a more complicated inhibitory mechanism involving more than one pathway for reversal of the tight complex. A slow-binding mechanism of inhibition has not been previously observed with peptidyldiazomethanes. Incorporation of [3H]Ac-Ala-Ala-Pro-diazomethane into prolyl endopeptidase was observed after denaturation of the inhibited complex. The peptide labelled with [3H]Ac-Ala-Ala-Pro-diazomethane was isolated and found to contain the active-site serine residue.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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