Solution parameters modulating DNA binding specificity of the restriction endonuclease EcoRV.

ABSTRACT: The DNA binding stringency of restriction endonucleases is crucial for their proper function. The X-ray structures of the specific and non-cognate complexes of the restriction nuclease EcoRV are considerably different suggesting significant differences in the hydration and binding free energies. Nonetheless, the majority of studies performed at pH 7.5, optimal for enzymatic activity, have found a < 10-fold difference between EcoRV binding constants to the specific and nonspecific sequences in the absence of divalent ions. We used a recently developed self-cleavage assay to measure EcoRV-DNA competitive binding and to evaluate the influence of water activity, pH and salt concentration on the binding stringency of the enzyme in the absence of divalent ions. We find the enzyme can readily dis

SUBMITTER: Sidorova NY 

PROVIDER: S-EPMC3156093 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature