Action on bovine alpha(s1)-casein of cardosins A and B, aspartic proteinases from the flowers of the cardoon Cynara cardunculus L

被引:50
作者
RamalhoSantos, M [1 ]
Verissimo, F [1 ]
Faro, C [1 ]
Pires, E [1 ]
机构
[1] UNIV COIMBRA, FAC CIENCIAS & TECNOL, DEPT BIOQUIM, P-3000 COIMBRA, PORTUGAL
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 1996年 / 1297卷 / 01期
关键词
cardosin; aspartic proteinase; proteolytic specificity; alpha(s1)-casein; chymosin; (Cynara cardunculus L);
D O I
10.1016/0167-4838(96)00103-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The cleavage of purified bovine alpha(s1)-casein separately by cardosin A and cardosin B, two distinct milk-clotting aspartic proteinases (APs) present in the stigmas of the plant Cynara cardunculus L., was studied. Casein digestion peptides were separated either by SDS-PAGE or by reverse-phase HPLC, and their N-terminal amino-acid sequences were subsequently determined by automated Edman degradation, thus identifying the cleavage sites. Results showed that both enzymes exert a similar but distinct action on bovine alpha(s1)-casein. In common they have the preference for the bond Phe(23)-Phe(24), and the cleavage of Trp(164)-Tyr(165) and Phe(153)-Tyr(154). Cardosin A also cleaves the bond Tyr(165)-Tyr(166), whereas Cardosin B cleaves an extra type of bond, Phe(150)-Arg(151), revealing a slightly broader specificity. A model for the action of both enzymes on bovine alpha(s1)-casein is proposed and discussed. In comparison with the reported action of chymosin on bovine alpha(s1)-casein, both cardosins proved to have a broader specificity towards this particular substrate due to a higher ability to cleave bonds between residues with large hydrophobic side-chains.
引用
收藏
页码:83 / 89
页数:7
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