Intramolecularly‐Quenched Fluorescent Peptides as Fluorogenic Substrates of Leucine Aminopeptidase and Inhibitors of Clostridial Aminopeptidase
- 1 March 1977
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 73 (2), 617-625
- https://doi.org/10.1111/j.1432-1033.1977.tb11357.x
Abstract
Fluorogenic oligopeptide derivatives of the type Lys(ABz)-X-ONBzl, where ABz is o-aminobenzoyl (anthraniloyl), X stands for Ala, Phe or Ala-Ala and ONBzl is p-nitrobenzyloxy, were synthesized and hydrolyzed by leucine aminopeptidase [swine kidney]. The hydrolysis is accompanied by an increase in fluorescence due to disruption of the intramolecular quenching of the fluorescent anthraniloyl moiety by the nitrobenzyl ester group. The spectral characteristics of the compounds are not consistent with an energy transfer mechanism according to Forster, therefore the quenching is assumed to be caused by a direct encounter between the quenching and the fluorescent groups. The change in fluorescence that accompanies the enzyme hydrolysis of the 1st peptide bond was used for quantitative measurement of the activity of leucine aminopeptidase and for the determination of some of its kinetic parameters. A bacterial aminopeptidase [EC 3.4.11] from Clostridium histolyticum that is very similar to leucine aminopeptidase in its substrate specificity did not hydrolyze the above peptide derivatives. The hydrolysis of leucine p-nitroanilide by this enzyme was inhibited by the 3 peptides and the corresponding inhibition constants were determined.This publication has 19 references indexed in Scilit:
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