Active human cytomegalovirus protease is a dimer

被引:84
作者
Darke, PL [1 ]
Cole, JL [1 ]
Waxman, L [1 ]
Hall, DL [1 ]
Sardana, MK [1 ]
Kuo, LC [1 ]
机构
[1] MERCK SHARP & DOHME RES LABS,DEPT BIOL CHEM,W POINT,PA 19486
关键词
D O I
10.1074/jbc.271.13.7445
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 quaternary state of the human cytomegalovirus (hCMV) protease has been analyzed in relation to its catalysis of peptide hydrolysis. Based on results obtained from steady state kinetics, size exclusion chromatography, and velocity sedimentation, the hCMV protease exists in a monomer-dimer equilibrium. Dimerization of the protease is enhanced by the presence of glycerol and high concentrations of enzyme. Isolation of monomeric and dimeric species eluted from a size exclusion column, followed by immediate assay, identifies the dimer as the active species. Activity measurements conducted with a range of enzyme concentrations are also consistent with a kinetic model in which only the dimeric hCMV protease is active. Using this model, the dissociation constant of the protease is 6.6 mu M in 10% glycerol and 0.55 mu M in 20% glycerol at 30 degrees C and pH 7.5.
引用
收藏
页码:7445 / 7449
页数:5
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