Conformational heterogeneity of Savinase from NMR, HDX-MS and X-ray diffraction analysis.

ABSTRACT:

Background

Several examples have emerged of enzymes where slow conformational changes are of key importance for function and where low populated conformations in the resting enzyme resemble the conformations of intermediate states in the catalytic process. Previous work on the subtilisin protease, Savinase, from Bacillus lentus by NMR spectroscopy suggested that this enzyme undergoes slow conformational dynamics around the substrate binding site. However, the functional importance of such dynamics is unknown.

Methods

Here we have probed the conformational heterogeneity in Savinase by following the temperature dependent chemical shift changes. In addition, we have measured changes in the local stability of the enzyme when the inhibitor phenylmethylsulfonyl fluoride is

SUBMITTER: Wu S 

PROVIDER: S-EPMC7323712 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature