Lytic protease can exist in two separately stable conformations with different His57 mobilities and catalytic activities

Lytic protease can exist in two separately stable conformations with different His57 mobilities and catalytic activities
Kristin Coffman Haddad, James L. Sudmeier, Daniel A. Bachovchin and William W. Bachovchin
January 25, 2005
PNAS
Lytic protease is a bacterial serine protease widely studied as a model system of enzyme catalysis. Here we report that lyophilization induces a structural change in the enzyme that is not reversed by redissolution in water. The structural change reduces the mobility of the active-site histidine residue and the catalytic activity of the enzyme. The application of mild pressure to solutions of the altered enzyme reverses the lyophilization-induced structural change and restores the mobility of the histidine residue and the enzyme's catalytic activity. This effect of lyophilization permits a unique opportunity for investigating the relationship between histidine ring dynamics and catalytic activity. The results demonstrate that His57 in resting enzymes is more mobile than previously thought, especially when protonated. The histidine motion and its correlation to enzyme activity lend support to the reaction-driven ring flip hypothesis.
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