Cloning and sequence analysis of cDNA for bovine carboxypeptidase E.
Carboxypeptidase E (enkephalin convertase) was first identified as the carboxypeptidase B-like enzyme involved in the biosynthesis of enkephalin in bovine adrenal chromaffin granules. A similar enzyme is present in many brain regions and in purified secretory granules from rat pituitary and rat insulinoma. Within the secretory granules, carboxypeptidase E (CPE) activity is found in both a soluble and a membrane-bound form, which differ slightly in relative molecular mass (Mr). Here, to investigate whether the CPE activities in the various tissues are produced from a single gene, purified CPE was partially sequenced and oligonucleotide probes were used to isolate a clone encoding CPE from a bovine pituitary complementary DNA library. This cDNA hybridizes to bovine pituitary poly(A)+ RNAs of approximately 3.3, 2.6 and 2.1 kilobases (kb), with the 3.3-kb messenger RNA the predominant species. The predicted amino-acid sequence of the cDNA clone contains the partially determined sequences of CPE, several pairs of basic amino acids and displays some homology with both carboxypeptidases A and B. Restriction analysis of bovine genomic DNA suggests only one gene for CPE. This is consistent with a broad role for CPE in the biosynthesis of many neuropeptides.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000235:genetics*; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA; D004262:DNA Restriction Enzymes; D009693:Nucleic Acid Hybridization; D010902:Pituitary Gland / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D004247:DNA; D004262:DNA Restriction Enzymes; D002268:Carboxypeptidases; D043423:Carboxypeptidase H