Facilitated Distortion of the DNA Site Enhances EcoRI Endonuclease-DNA Recognition

We have measured the binding of EcoRI endonuclease to a complete set of purine-base analogue sites, each of which deletes one functional group that forms a hydrogen bond with the endonuclease in the canonical GAATTC complex. For five of six functional group deletions, the observed penalty in binding...

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Published in:Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 16; pp. 7548 - 7552
Main Authors: Lesser, David R., Kurpiewski, Michael R., Waters, Timothy, Connolly, Bernard A., Jen-Jacobson, Linda
Format: Journal Article
Language:English
Published: Washington, DC National Academy of Sciences of the United States of America 15-08-1993
National Acad Sciences
National Academy of Sciences
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Summary:We have measured the binding of EcoRI endonuclease to a complete set of purine-base analogue sites, each of which deletes one functional group that forms a hydrogen bond with the endonuclease in the canonical GAATTC complex. For five of six functional group deletions, the observed penalty in binding free energy is +1.3 to +1.7 kcal/mol. For two of these cases (replacement of adenine N7 with carbon) a single protein-base hydrogen bond is removed without deleting an interstrand Watson-Crick hydrogen bond or causing structural "adaptation" in the complex. This observation establishes that the incremental energetic contribution of one protein-base hydrogen bond is about -1.5 kcal/mol. By contrast, deletion of the N6-amino group of the inner adenine in the site improves binding by -1.0 kcal/mol because the penalty for deleting a protein-base hydrogen bond is outweighed by facilitation of the required DNA distortion ("kinking") in the complex. This result provides direct evidence that the energetic cost of distorting a DNA site can make an unfavorable contribution to protein-DNA binding.
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ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.90.16.7548