title: Fluorogenic peptide substrates for assay of aspartyl proteinases.
abstract: Via a combination of chemical and enzymatic synthesis, new hexapeptide substrates convenient for use in activity assessment of several aspartyl proteinases--porcine pepsin, human pepsin, gastricsin, and cathepsin D--were prepared. These peptide derivatives, o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Ala-p-nitroanilide and N-(o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Ala)-N'-2,4-dinitrophenyl ethylenediamine, contain a fluorescent o-aminobenzoyl moiety as well as p-nitroaniline or N-2,4-dinitrophenyl ethylenediamine--the groups that cause fluorescence quenching. Aspartyl proteinases hydrolyze the Phe-Phe peptide bond in the substrates, which diminishes quenching due to separation of the fluorescent and quenching moieties and leads to an increase in the fluorescence intensity of o-aminobenzoyl residue. Abz-Ala-Ala-Phe-Phe-Ala-Ala-Ded, being fairly well hydrolyzed by HIV proteinase, might be used for assay of this enzyme.
authors.affiliation: Department of Chemistry, Moscow State University, Russia.
authors.forename: I Y
authors.initials: IY
authors.lastname: Filippova
authors.forename: E N
authors.initials: EN
authors.lastname: Lysogorskaya
authors.forename: V V
authors.initials: VV
authors.lastname: Anisimova
authors.forename: L I
authors.initials: LI
authors.lastname: Suvorov
authors.forename: E S
authors.initials: ES
authors.lastname: Oksenoit
authors.forename: V M
authors.initials: VM
authors.lastname: Stepanov
mesh_terms: D000595:Amino Acid Sequence; D062365:Aminobenzoates / Q000737:chemistry*; D000818:Animals; D002402:Cathepsin D / Q000032:analysis*; D005470:Fluorometry; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009842:Oligopeptides / Q000737:chemistry*; D010434:Pepsin A / Q000032:analysis*; D013552:Swine
chemical_list: C101257:2-aminobenzoyl-alanyl-alanyl-phenylalanyl-phenylalanyl-alanyl-alanyl-4-nitroanilide; D062365:Aminobenzoates; C101258:N-(2-aminobenzoyl-alanyl-alanyl-phenylalanyl-phenylalanyl-alanyl-alanyl)-N'-2,4-dinitrophenylethylenediamine; D009842:Oligopeptides; D010434:Pepsin A; C020557:gastricsin; D002402:Cathepsin D