Synthesis and protease-catalyzed hydrolysis of a novel hydrazinopeptide.
In order to explore the potentiality of hydrazinopeptides as protease inhibitors, the resistance of the hydrazinopeptidic bond toward proteolysis was investigated. To this end, the novel hydrazinohexapeptide Z-Ala2-Pro-Val-hIle-Leu-OMe (1), where hIle represents hydrazinoisoleucine, was designed and synthesized together with the parent peptide Z-Ala2-Pro-Val-Ile-Leu-OMe (2). The interactions of 1 and 2 with human leukocyte elastase (HLE) and porcine pancreatic elastase (PPE) were analyzed comparatively. We observed that 1 behaved as a substrate for both elastases, without the formation of a stable acyl-enzyme as in the case of azapeptides. Compounds 1 and 2 were cleaved at the same site (-Val-parallel-NH-) with a slight delay of hydrolysis for 1 compared to 2 (kcat/KM for 1 vs. 2 decreased by a factor of 2.7 for the HLE-catalyzed hydrolysis at pH 8.0 and 25 degrees C). The presence of the hydrazinopeptide bond (-CONHNH-) in 1 reduced by a factor of 4.7 the apparent enzyme affinity without abolishing it. These results indicate that suitably designed hydrazinopeptides may represent interesting targets in the search for protease resisting pseudopeptides.
Amour
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Department of Cellular and Supramolecular Biology, Institute Jacques Monod, University of Paris VII, France.
Collet
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Dubar
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Reboud-Ravaux
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D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D006801:Humans; D006834:Hydrazines / Q000138:chemical synthesis* / Q000378:metabolism* / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D019272:Leukocyte Elastase; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000378:metabolism* / Q000494:pharmacology; D010179:Pancreas / Q000201:enzymology; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D013552:Swine
D006834:Hydrazines; D009842:Oligopeptides; D011480:Protease Inhibitors; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase