1.[p-Nitroanilides of amino acids and peptides and fluorescence peptide with inner fluorescence quenching as substrates for cathepsins H, B, D and high molecular weight aspartic peptidase in the brain].
Azarian AV, Agatian GL, Galoian AA. Biokhimiia. 1987 Dec;52(12):2033-7.
p-Nitroanilides of amino acids and peptides were used to study the specificity of cathepsins H and B from human and bovine brain, respectively. The specific activity of cathepsin H decreased in the following order: Arg-pNa greater than or equal to Leu-pNa greater than Ala-pNa greater than or equal to Phe-pNa greater than Pro-pNa greater than Glu-pNa; Arg-pNa was split by the enzyme 12 times as fast as Bz-Arg-pNa. Among other oligopeptide p-nitroanilides tested (Ala-Ala, Ala-Leu, Ala-Ala-Ala, Ala-Ala-Leu, Gly-Gly-Leu, Gly-Gly-Phe, Gly-Leu-Phe, pGlu-Phe-Leu, pGlu-Phe-Ala, pGlu-Phe), PGlu-Phe-Leu and pGlu-Phe-Ala appeared to be the best substrates for cathepsin B; Km for hydrolysis were 0.1 mM and 0.165 mM, respectively, kcat were 5.1 and 8.3 s-1, respectively. A comparative study of substrate specificity of cathepsin D and high molecular weight aspartic peptidase with the use of fluorescent substrate with inner fluorescence quenching, Abz-Ala-Ala-Phe-Phe-pNa, revealed that both peptidases hydrolyzed the single bond between two phenylalanine residues, resulting in the increase of fluorescence (4.
