Abstract
The primary structure of malt carboxypeptidase I has been determined. The enzyme is composed of two peptide chains, an A- and a B-chain, linked by disulphide bridges. Fragments were obtained by chemical cleavages with either cyanogen bromide or hydroxylamine and by enzymatic cleavages with either trypsin, S. aureus V8 protease or proline specific endopeptidase (E.C. 3.4.21.26), sequenced and aligned to give the total sequence of the two chains. The A- and B-chains contain 266 and 148 amino acid residues, respectively. Glycosylated asparagines are found in positions 118 and 232 of the A-chain and position 56 of the B-chain. Alignment of the sequence of the A-chain with the N-terminal part of carboxypeptidase Y revealed 30% homology. Similarly, the B-chain showed 27% homology with the C-terminal part of carboxypeptidase Y. No homology was observed with other proteins by a computer search of a sequence data base provided by the National Biomedical Research Foundation. A region of the A-chain was found to be identical to the region around the essential seryl residue in position 146 of carboxypeptidase Y.
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Abbreviations
- CABS-Sepharose:
-
[N-(ε-aminocaproyl)-p-amino-benzyl]succiny]-Sepharose 4B
- DPCC:
-
diphenylcarbamyl chloride
- EDTA:
-
ethylenediaminetetraacetic acid, disodium salt
- HPLC:
-
high pressure liquid chromatography
- PTH:
-
phenylthiohydantoin
- TFA:
-
trifluoroacetic acid
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The binding site notations for the enzyme is that ofSchechter andBerger (17). Accordingly, the binding site for the C-terminal amino acid residue of the substrate is denoted S′1 and those for the amino acid residues in the amino-terminal direction away from the scissile bond are denoted S1, S2,… Sn. Amino acid residues in the substrate are referred to as P1, P2,…, Pa and P′1 in correspondence with the binding site.
Accepted byE. Lund
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Sørensen, S.B., Breddam, K. & Svendsen, I. Primary structure of carboxypeptidase I from malted barley. Carlsberg Res. Commun. 51, 475–485 (1986). https://doi.org/10.1007/BF02906889
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DOI: https://doi.org/10.1007/BF02906889