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Nickel-mediated sequence-specific oxidative cleavage of DNA by a designed metalloprotein

Mack, David P. and Dervan, Peter B. (1990) Nickel-mediated sequence-specific oxidative cleavage of DNA by a designed metalloprotein. Journal of the American Chemical Society, 112 (11). pp. 4604-4606. ISSN 0002-7863. doi:10.1021/ja00167a102.

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We recently designed a sequence-specific DNA-cleaving metalloprotein consisting wholly of naturally occurring α-amino acids. The tripeptide copper-binding domain, Gly-Gly-His (GGH), was attached to the NH2-terminus of the DNA-binding domain of Hin recombinase (residues 139-1 90) to afford a new 55-residue protein, GGH(Hin 139-1 90). This protein is capable of binding DNA at four 13 base pair sites (termed hixL and secondary). CuDGGH(Hin139-190) in the presence of excess hydrogen peroxide and sodium ascorbate cleaves DNA predominantly at one of the four Hin binding sites. We report here that, in the presence of Ni(OAc), and monoperoxyphthalic acid, the sequence specificity and efficiency of the DNA cleavage by GGH(Hin139-190) are remarkably altered. The nickel-mediated DNA cleavage process occurs at all four binding sites, is more rapid and efficient, and is chemically activated by 1 equiv of an oxygen atom donor. At the hixL site, cleavage occurs predominantly at a single deoxyribose position on one strand of each binding site, indicating a nondiffusible oxidizing species.

Item Type:Article
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Dervan, Peter B.0000-0001-8852-7306
Additional Information:© 1990 American Chemical Society. Received February 13, 1990. Acknowledgment. We are grateful for support from the DARPA University Research Initiative Program, Merck and for a National Research Service Award to D.P.M. We thank M. Greenberg for helpful discussions.
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)UNSPECIFIED
NIH Predoctoral FellowshipUNSPECIFIED
Issue or Number:11
Record Number:CaltechAUTHORS:20160510-151735708
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Official Citation:Nickel-mediated sequence-specific oxidative cleavage of DNA by a designed metalloprotein David P. Mack and Peter B. Dervan Journal of the American Chemical Society 1990 112 (11), 4604-4606 DOI: 10.1021/ja00167a102
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:66959
Deposited By: Victoria Brennan
Deposited On:19 May 2016 18:41
Last Modified:11 Nov 2021 00:24

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