Fluorogenic Peptide Substrates for Assay of Aspartyl Proteinases

✍ Scribed by Irina Yu. Filippova; Elena N. Lysogorskaya; Veronika V. Anisimova; Leonid I. Suvorov; Elena S. Oksenoit; Valentin M. Stepanov

• Publisher: Elsevier Science
• Year: 1996
• Tongue: English
• Weight: 121 KB
• Volume: 234
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1996.0062

✦ Synopsis

should possess more or less extended peptide chain. As Via a combination of chemical and enzymatic syn-a rule, S 1 and S 1 subsites accommodate the side chains thesis, new hexapeptide substrates convenient for of hydrophobic amino acids-the feature often used to use in activity assessment of several aspartyl proconstruct specific substrates for these proteinases.

teinases -porcine pepsin, human pepsin, gastricsin,

A number of synthetic substrates have been sugand cathepsin D -were prepared. These peptide degested for aspartyl proteinases, including those conrivatives, o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Alataining p-nitrophenylalanine residues at the susceptip-nitroanilide and N-(o-aminobenzoyl-Ala-Ala-Pheble peptide bond (2-4). In addition to these Phe-Ala-Ala)-N -2,4-dinitrophenyl ethylenediamine, chromogenic substrates, the application of which is contain a fluorescent o-aminobenzoyl moiety as well handicapped by a relatively low molar extinction increas p-nitroaniline or N-2,4-dinitrophenyl ethylenediment that accompanies the cleavage of the peptide amine -the groups that cause fluorescence quenchbond, fluorogenic substrates have been suggested for ing. Aspartyl proteinases hydrolyze the Phe-Phe use in assessing aspartyl proteinases. These compeptide bond in the substrates, which diminishes pounds contain both a fluorophore and a quenching quenching due to separation of the fluorescent and moiety within the same molecule, which leads to interquenching moieties and leads to an increase in the nal fluorescence quenching. After the rupture of the fluorescence intensity of o-aminobenzoyl residue. peptide chain, the efficiency of the quenching decreases Abz-Ala-Ala-Phe-Phe-Ala-Ala-Ded, being fairly well substantially, resulting in a more or less sharp increase hydrolyzed by HIV proteinase, might be used for in fluorescence emission and thus allowing a fast and assay of this enzyme.