Cloning and expression of human carboxypeptidase Z, a novel metallocarboxypeptidase

Lixin Song, Lloyd D. Fricker

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

A novel cDNA, designated carboxypeptidase Z (CPZ), was identified based on its homology to known metallocarboxypeptidases. Northern blot analysis shows bands of 2.1 and/or 2.6 kilobases in all tissues examined. The major form of CPZ mRNA in human salivary gland encodes a protein with an open reading frame of 641 amino acids. In addition, three variants were found that presumably arise due to alternative intron splicing. The 641-amino acid protein contains an 18-residue signal peptide-like sequence, a 120-residue region that shows 23-29% amino acid identity with a Cys-rich domain found in frizzled proteins and in type XVIII collagen, and then a 390-residue carboxypeptidase domain with 49% amino acid identity to carboxypeptidases E and N.The 641-amino acid form of CPZ expressed in the baculovirus system cleaves 5-dimethylaminonaphthalene-1-sulfonyl (dansyl)-Phe-Ala-Arg, although the level of enzyme activity was approximately 10-fold lower than either carboxypeptidase E or D expressed using the same vital system. The CPZ activity is more active at neutral pH than at pH 5.5 and is inhibited by active site-directed inhibitors of metallocarboxypeptidases. In summary, CPZ is a novel metallocarboxypeptidase that is active toward substrates with C- terminal basic amino acids.

Original languageEnglish (US)
Pages (from-to)10543-10550
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number16
DOIs
StatePublished - Apr 18 1997

Fingerprint

Cloning
Organism Cloning
Amino Acids
Carboxypeptidase H
Collagen Type XVIII
Lysine Carboxypeptidase
Frizzled Receptors
Cathepsin A
Carboxypeptidases
Basic Amino Acids
Baculoviridae
Alternative Splicing
Enzyme activity
Protein Sorting Signals
Salivary Glands
Northern Blotting
Introns
Open Reading Frames
Catalytic Domain
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cloning and expression of human carboxypeptidase Z, a novel metallocarboxypeptidase. / Song, Lixin; Fricker, Lloyd D.

In: Journal of Biological Chemistry, Vol. 272, No. 16, 18.04.1997, p. 10543-10550.

Research output: Contribution to journalArticle

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