Molecular mechanism for dimerization to regulate the catalytic activity of human cytomegalovirus protease

被引:48
作者
Batra, R [1 ]
Khayat, R [1 ]
Tong, L [1 ]
机构
[1] Columbia Univ, Dept Biol Sci, New York, NY 10027 USA
基金
美国国家卫生研究院;
关键词
D O I
10.1038/nsb0901-810
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
Biochemical studies indicate that dimerization is required for the catalytic activity of herpesvirus proteases, whereas structural studies show a complete active site in each monomer, away from the dimer interface. Here we report kinetic, biophysical and crystallographic characterizations of structure-based mutants in the dimer interface of human cytomegalovirus (HCMV) protease. Such mutations can produce a 1,700-fold reduction in the k(cat) while having minimal effects on the K-m. Dinner stability is not affected by these mutations, suggesting that dimerization itself is insufficient for activity. There are large changes in monomer conformation and dimer organization of the apo S225Y mutant enzyme. However, binding of an activated peptidomimetic inhibitor induced a conformation remarkably similar to the wild type protease. our studies suggest that appropriate dimer formation may be required to indirectly stabilize the protease oxyanion hole, revealing a novel mechanism for dimerization to regulate enzyme activity.
引用
收藏
页码:810 / 817
页数:8
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