Trypsin-Chymotrypsin Inhibitors from Vigna mungo Seeds

被引:20
作者
Cheung, Allen H. K. [1 ]
Wong, Jack H. [1 ]
Ng, T. B. [1 ]
机构
[1] Chinese Univ Hong Kong, Fac Med, Dept Biochem, Shatin, Hong Kong, Peoples R China
关键词
HIV-1; REVERSE-TRANSCRIPTASE; BEAN PROTEASE INHIBITOR; CORONAVIRUS MAIN PROTEINASE; MOMORDICA-COCHINCHINENSIS SEEDS; AMINO-ACID-SEQUENCES; TERTIARY STRUCTURE; CHEMICAL-MODIFICATION; POTENT INHIBITOR; CLEAVAGE SITES; DRUG DESIGN;
D O I
10.2174/092986609787601714
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
Three trypsin-chymotrypsin inhibitors were isolated from seeds of the black gram (Vigna mungo) with a procedure that entailed cation exchange chromatography on SP-Sepharose, anion exchange chromatography on Q-Sepharose, ion exchange chromatography by fast protein liquid chromatography (FPLC) on Mono Q and Mono S, and gel filtration by FPLC on Superdex 75. Two of the trypsin-chymotrypsin inhibitors were adsorbed on the first four types of chromatographic media. All three inhibitors have a molecular mass of 16 kDa as judged by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The trypsin inhibitory activity of the inhibitors was attenuated in the presence of the reducing agent dithiothreitol. The remaining inhibitor was unadsorbed on SP-Sepharose but adsorbed on Q-Sepharose, Mono Q and Mono S. The protease inhibitors did not exert any inhibitory effect on hepatoma (Hep G2) and breast cancer (MCF 7) cells or antifungal action toward Botrytis cinerea, Fusarium oxysporum and Mycosphaerella arachidicola. Two of the inhibitors slightly inhibited the activity of HIV-1 reverse transcriptase, with an IC50 in the millimolar range.
引用
收藏
页码:277 / 284
页数:8
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