title: The A- and B-chains of carboxypeptidase I from germinated barley originate from a single precursor polypeptide.
abstract: Carboxypeptidase I from germinated barley (Hordeum vulgare) grain consists of two peptide chains linked by disulfides; the A- and B-chains contain 266 and 148 amino acid residues, respectively (Sorensen, S. B., Breddam, K., and Svendsen, I. (1986) Carlsberg Res. Commun. 51, 475-485). A cDNA library prepared from mRNA isolated from scutella of 2-day germinated barley has now been screened with a mixed oligonucleotide encoding a peptide fragment of the A-chain. Nucleotide sequence analysis of a 1443-nucleotide pair cDNA clone revealed that both chains of the enzyme are translated from a single mRNA. The coding region of the A-chain is located at the 5'-end of the cDNA and is separated from the B-chain coding region by a 165-nucleotide pair linking region. The B-chain coding region is followed by a stop codon, a 187-nucleotide pair 3'-untranslated sequence, and a short polyadenylic acid tail. The results indicate that the A- and B-chains of barley carboxypeptidase I arise by endoproteolytic excision of a 55-residue linker peptide from a single precursor polypeptide chain. The putative linker peptide is rich in proline, lysine, and arginine residues, has an apparent pI of 11.9, and appears to be excised by cleavage of peptide bonds on the COOH-terminal side of serine residues.
authors.affiliation: Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia.
authors.forename: N P
authors.initials: NP
authors.lastname: Doan
authors.forename: G B
authors.initials: GB
authors.lastname: Fincher
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002268:Carboxypeptidases / Q000032:analysis*; D004247:DNA / Q000032:analysis; D001467:Hordeum; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010940:Plant Proteins; D010944:Plants / Q000201:enzymology*
chemical_list: D010940:Plant Proteins; D004247:DNA; D002268:Carboxypeptidases; C056823:CP-MI protein, Hordeum vulgare