title: Chloride-insensitive, glycine-phenylalanine-naphthylamide hydrolysis at neutral pH in human skin fibroblasts.
abstract: Crude lysosomal preparations from a cultured human skin fibroblast line were found to contain significant levels of a neutral pH hydrolase activity towards glycine--phenylalanine--beta-naphthylamide (NA), a substrate normally used for the assay of lysosomal dipeptidyl aminopeptidase I. However, the activity was chloride ion insensitive, nonlatent, and inhibitable by cationic detergents and amino acids. Assays of substrate selectivity, relative substrate affinity, pH and anion and cation sensitivity indicated the activity to be distinct from dipeptidyl aminopeptidases I (chloride-dependent hydrolysis of Pro-Phe-, Gly-Phe-, Gly-Arg-, and Pro-Arg-NA's at acid pH), II (Lys-Ala-NA hydrolysis), III (Arg-Arg-NA hydrolysis), and IV (Gly-Pro-NA hydrolysis). The lysosomal preparations also contained significant activity towards several amino acid--naphthylamides, notably Arg-NA. Only dipepidyl aminopeptidase I activity showed sensitivity to chloride anions, both dipeptidyl aminopeptidases I and II showed substantial latency, and none of the activities displayed a significant metal cation dependent.
mesh_terms: D000328:Adult; D000838:Anions; D002412:Cations; D002460:Cell Line; D002712:Chlorides / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology*; D005998:Glycine / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008670:Metals / Q000494:pharmacology; D009281:Naphthalenes / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism*; D012867:Skin / Q000201:enzymology
chemical_list: D000838:Anions; D002412:Cations; D002712:Chlorides; D003902:Detergents; D008670:Metals; D009281:Naphthalenes; D010649:Phenylalanine; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D005998:Glycine