One- and two-metal ion catalysis: global single-turnover kinetic analysis of the PvuII endonuclease mechanism.

ABSTRACT: Ester hydrolysis is one of the most ubiquitous reactions in biochemistry. Many of these reactions rely on metal ions for various mechanistic steps. A large number of metal-dependent nucleases have been crystallized with two metal ions in their active sites. In spite of an ongoing discussion about the roles of these metal ions in nucleic acid hydrolysis, there are very few studies which examine this issue using the native cofactor Mg(II) and global fitting of reaction progress curves. As part of a comprehensive study of the representative homodimeric PvuII endonuclease, we have collected single-turnover DNA cleavage data as a function of Mg(II) concentration and globally fit these data to a number of models which test various aspects of the metallonuclease mechanism. DNA association rate co

SUBMITTER: Xie F 

PROVIDER: S-EPMC2728687 | biostudies-literature | 2008 Nov

REPOSITORIES: biostudies-literature