Sequencing and cloning of human prolylcarboxypeptidase (angiotensinase C). Similarity to both serine carboxypeptidase and prolylendopeptidase families.
Prolylcarboxypeptidase, a lysosomal serine carboxypeptidase, cleaves COOH-terminal amino acids linked to proline, as in angiotensin II and III and [des-Arg9] bradykinin. About 25% of the enzyme protein was sequenced, and the complete sequence was deduced from its human kidney cDNA. The cDNA insert contained an open reading frame of 1488 base pairs coding for a protein of 496 residues. The authentic NH2-terminal sequence matched the deduced protein sequence starting with residue 46, suggesting the presence of both a signal and propeptide. The mature enzyme (451 residues) has a calculated M(r) = 51,043, whereas the M(r) of the purified glycoprotein is 58,000, indicating 12% carbohydrate. The overall sequence identity with serine peptidases is low (10-18%), but sequences around residues of the putative catalytic triad (Ser134, Asp333, His411) are similar (30-67%) to both the serine carboxypeptidases (e.g. deamidase or lysosomal protective protein, yeast carboxypeptidase Y, and KEX1 gene product) and the prolylendopeptidase family. Thus, prolylcarboxypeptidase links these two families, suggesting an evolutionary relationship. It is inhibited (Ki = 2.6 x 10(-7) M) by benzyloxycarbonyl-Pro-prolinal, a specific inhibitor of prolylendopeptidase, another angiotensin metabolizing enzyme. Prolylcarboxypeptidase contains serine or threonine residues repeated as the 26th residue 7 out of 9 times, with identical or similar amino acids in other positions in the repeats. The KEX1 gene product contains a similar motif, with serine or threonine as every 27th residue. The importance of prolylcarboxypeptidase is strongly suggested by its presence in various organs and cells and by the substrates it cleaves.
Tan
F
F
Department of Pharmacology, University of Illinois College of Medicine, Chicago 60612.
Morris
P W
PW
Skidgel
R A
RA
Erdös
E G
EG
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004247:DNA; D006801:Humans; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics
D004247:DNA; D002268:Carboxypeptidases; C022448:lysosomal Pro-X carboxypeptidase; C022633:serine carboxypeptidase; D012697:Serine Endopeptidases