Abstract
A serine carboxypeptidase isolated from malted barley by affinity chromatography was termed malt carboxypeptidase II to distinguish it from another malt carboxypeptidase previously described (Carlsberg Res. Commun. 48, 217–230 (1983)), henceforth called malt carboxypeptidase I. Our nomenclature is in agreement with the nomenclature formerly suggested byMikola. Malt carboxypeptidase II has a molecular weight of 110,000–120,000. It appears to be a dimer where each monomer is composed of two peptide chains linked by disulfide bridges: one monomer contains an A-chain (34,000) and a B-chain (27,000), the other an A-chain and a C-chain (24,000). The enzyme contains 28 residues of glucosamine and 15% neutral sugar. The N-terminal sequence of the A-chain was NH2-Ala-Gly-Gly-His-Ala-Ala-Asp-Arg-Ile-Val- while the B- and C-chains appeared to be N-terminally blocked. The amino acid compositions of the B- and C-chains were identical suggesting that their different molecular weights are due to different contents of carbohydrate.
Malt carboxypeptidase II is inhibited by diisopropyl phosphorofluoridate and by Hg++. It exhibits a strong preference for substrates containing Lys and Arg as C-terminal amino acid residues but it also hydrolyses substrates with hydrophobic amino acid residues in this position.
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Abbreviations
- BS:
-
benzyl succinic acid
- Bz:
-
benzoyl
- CABS-Sepharose:
-
[N-(ε-aminocaproyl)-p-amino-benzyl]succinyl-Sepharose 4B
- DFP:
-
diisopropyl phosphorofluoridate
- EDTA:
-
ethylenediamine tetraacetic acid, sodium salt
- FA:
-
furylacryloyl
- Hepes:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- HPLC:
-
high performance liquid chromatography
- Mes:
-
2-(N-morpholino) ethane sulfonic acid
- p-HMB:
-
parahydroxymercuribenzoate
- SDS:
-
sodium dodecyl sulfate
- Tris:
-
tris(hydroxy methyl)aminomethane
- Z:
-
carbobenzoxy
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Breddam, K., Sørensen, S.B. & Ottesen, M. Isolation of carboxypeptidase II from malted barley by affinity chromatography. Carlsberg Res. Commun. 50, 199–209 (1985). https://doi.org/10.1007/BF02907146
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DOI: https://doi.org/10.1007/BF02907146