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Published July 25, 1982 | Published
Journal Article Open

The presence of zinc in the restriction enzyme Eco RI

Abstract

We have determined that the restriction endonuclease Eco RI contains 1.0 ± 0.1 eq of zinc/monomeric enzyme. DNA cleavage by Eco RI is inhibited by ortho-phenanthroline after preincubation of the enzyme with the chelating agent. A similar inhibition by the nonchelating meta-phenanthroline is not seen. The sensitivity of the inhibition by the neutral ligand ortho-phenanthroline to preincubation is consistent with the tightly bound and inaccessible nature of the metal site. Extensive dialysis against the ortho-phenanthroline inhibitor leads to the release of the bound metal with the concomitant loss of enzyme activity. The tightly bound Zn^(2+) cation, then, appears to be necessary for enzyme function. The finding of zinc in Eco RI further illustrates the ubiquity of Zn^(2+) to DNA-protein complexes.

Additional Information

© 1982 American Society for Biochemistry and Molecular Biology. Received for publication, February 9, 1982. This work was supported by the American Chemical Society Petroleum Research Fund and the City University of New York PSCBHE Research Award Program. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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August 19, 2023
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