ACTIVATION OF THE HERPES-SIMPLEX VIRUS TYPE-1 PROTEASE

被引:58
作者
HALL, DL [1 ]
DARKE, PL [1 ]
机构
[1] MERCK SHARP & DOHME LTD,RES LABS,DEPT BIOL CHEM,W POINT,PA 19486
关键词
D O I
10.1074/jbc.270.39.22697
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 catalytic efficiency of the mature HSV-1 protease has been examined as a function of solvent composition, With the peptide substrate HTYLQASEKFKMWG-amide, the specificity constant (k(cat)/K-m) at pH 7.5 for cleavage is 5.2 M(-1) s(-1). This value increases to 38 Rr(-1) s(-1) when 25% glycerol is present in the reaction mixture. It was found that glycerol activation is but one case of the general phenomenon of HSV-1 protease activation by kosmotropes, or water structure-forming cosolvents. For example, an 860-fold increase in the protease activity (k(cat)/K-m = 4500 M(-1) s(-1)) occurs in the presence of 0.8 M sodium citrate. Similarly, the presence of 0.8 M sodium phosphate activates the catalytic efficiency by 420-fold (k(cat)/K-m = 2200 M(-1) s(-1)). The extent of HSV-1 protease activation by various anions correlates with the Hofmeister series. Both the susceptibility to proteolysis by trypsin and the protein fluorescence spectra of the HSV-1 protease change in the presence of activating solvents, suggesting a conformational change accompanying activation.
引用
收藏
页码:22697 / 22700
页数:4
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