title: Cloning and sequence analysis of cDNA encoding rat carboxypeptidase D.
abstract: Carboxypeptidase D (CPD) is a recently described 180-kD enzyme with carboxypeptidase E-like enzymatic properties. CPD has been proposed to be present in the secretory pathway and to contribute to peptide hormone processing in the Cpe(fat)/Cpe(fat) mouse, which lacks functional CPE. Sequence analysis of cDNA clones encoding rat CPD show the protein to contain an amino-terminal signal peptide, three carboxypeptidase-like domains, a putative transmembrane domain, and a 60-amino-acid cytoplasmic tail. Whereas active site, substrate-binding, and metal-binding residues of other metallocarboxypeptidases are conserved in the first two domains of CPD, several of the critical residues are not conserved in the third domain; this third domain is not predicted to form an active carboxypeptidase. The overall homology between rat CPD and the duck homolog gp180 is high, with 75% amino acid identity. The three carboxypeptidase domains show 66%, 83%, and 82% amino acid identity between rat CPD and duck gp180. Homology is also high in the transmembrane domain (86%) and in the cytoplasmic tail (97%). The mouse Cpd gene maps to the medial portion of chromosome 11, approximately 45.5 cM distal to the centromere. Northern blot analysis of CPD mRNA shows major bands of approximately 8 and 4 kb in many rat tissues, and additional species ranging from 1.4 to 5 kb that are expressed in some tissues or cell lines. CPD mRNA is detectable in most tissues examined, and is most abundant in hippocampus, spinal cord, atrium of the heart, colon, testis, and ovaries. In situ hybridization of CPD mRNA shows a distribution in many cells in rat brain and other tissues, with high levels in hippocampus, olfactory bulb, and the intermediate pituitary. The broad distribution is consistent with a role for CPD in the processing of many peptides and proteins that transit the secretory pathway.
authors.affiliation: Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: X
authors.initials: X
authors.lastname: Xin
authors.forename: O
authors.initials: O
authors.lastname: Varlamov
authors.forename: R
authors.initials: R
authors.lastname: Day
authors.forename: W
authors.initials: W
authors.lastname: Dong
authors.forename: M M
authors.initials: MM
authors.lastname: Bridgett
authors.forename: E H
authors.initials: EH
authors.lastname: Leiter
authors.forename: L D
authors.initials: LD
authors.lastname: Fricker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002268:Carboxypeptidases / Q000235:genetics*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003593:Cytoplasm / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics*; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D021382:Protein Sorting Signals / Q000235:genetics; D011506:Proteins; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D021382:Protein Sorting Signals; D011506:Proteins; D012333:RNA, Messenger; D002268:Carboxypeptidases; C094315:metallocarboxypeptidase D