title: Active site of tripeptidyl peptidase II from human erythrocytes is of the subtilisin type.
abstract: The present report presents evidence that the amino acid sequence around the serine of the active site of human tripeptidyl peptidase II is of the subtilisin type. The enzyme from human erythrocytes was covalently labeled at its active site with [3H]diisopropyl fluorophosphate, and the protein was subsequently reduced, alkylated, and digested with trypsin. The labeled tryptic peptides were purified by gel filtration and repeated reversed-phase HPLC, and their amino-terminal sequences were determined. Residue 9 contained the radioactive label and was, therefore, considered to be the active serine residue. The primary structure of the part of the active site (residues 1-10) containing this residue was concluded to be Xaa-Thr-Gln-Leu-Met-Asx-Gly-Thr-Ser-Met. This amino acid sequence is homologous to the sequence surrounding the active serine of the microbial peptidases subtilisin and thermitase. These data demonstrate that human tripeptidyl peptidase II represents a potentially distinct class of human peptidases and raise the question of an evolutionary relationship between the active site of a mammalian peptidase and that of the subtilisin family of serine peptidases.
authors.affiliation: Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
authors.forename: B
authors.initials: B
authors.lastname: Tomkinson
authors.forename: C
authors.initials: C
authors.lastname: Wernstedt
authors.forename: U
authors.initials: U
authors.lastname: Hellman
authors.forename: O
authors.initials: O
authors.lastname: Zetterqvist
mesh_terms: D000595:Amino Acid Sequence; D000626:Aminopeptidases; D001665:Binding Sites; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000097:blood*; D013381:Subtilisins; D014357:Trypsin
chemical_list: D010446:Peptide Fragments; D010447:Peptide Hydrolases; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; C048001:tripeptidyl-peptidase 2; D012697:Serine Endopeptidases; D013381:Subtilisins; D014357:Trypsin