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Synthesis of a hybrid protein containing the iron-binding ligand of bleomycin and the DNA-binding domain of Hin

Oakley, Martha G. and Turnbull, Kenneth D. and Dervan, Peter B. (1994) Synthesis of a hybrid protein containing the iron-binding ligand of bleomycin and the DNA-binding domain of Hin. Bioconjugate Chemistry, 5 (3). pp. 242-247. ISSN 1043-1802.

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The iron-binding and oxygen-activating domain of the natural product bleomycin [pyrimidoblamic acid-beta-hydroxy-L-histidine (PBA-beta-OH-His)] was attached to the NH_2 terminus of the DNA binding domain of Hin recombinase (residues 139-190). This hybrid 54-residue protein PBA-beta-OH-His-Hin-(139-190) binds specifically to DNA at four distinct Hin binding sites with affinities comparable to those of the unmodified Hin(139-190). In the presence of dithiothreitol, Fe(II).PBA-beta-OH-His-Hin-(139-190) cleaves DNA with specificity remarkably similar to that of Fe(II).EDTA-Hin(139-190). Analysis of the cleavage patterns suggests that site-specific DNA cleavage is mediated by a localized diffusible species, in contrast with cleavage by bleomycin, which occurs through a nondiffusible oxidant. This has implications for the design of second-generation artificial sequence specific DNA cleaving proteins and defines limitations in current efforts to create atom-specific chemistry on DNA.

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Dervan, Peter B.0000-0001-8852-7306
Additional Information:© 1994 American Chemical Society. Received February 16, 1994. Abstract published in Advance ACS Abstracts, April 15, 1994. This work was supported by the National Institutes of Health (GM-276811, a National Science Foundation predoctoral fellowship, a National Institutes of Health predoctoral traineeship, and a Glaxo Graduate Student Fellowship to M.G.O. and an American Cancer Society postdoctoral fellowship to K.D.T. We thank G. M. Hathaway (University of California, Riverside) for protein mass spectral analysis.
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NSF Predoctoral FellowshipUNSPECIFIED
NIH Predoctoral FellowshipUNSPECIFIED
American Cancer SocietyUNSPECIFIED
Issue or Number:3
Record Number:CaltechAUTHORS:20160510-133747674
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Official Citation:Synthesis of a Hybrid Protein Containing the Iron-Binding Ligand of Bleomycin and the DNA-Binding Domain of Hin Martha G. Oakley, Kenneth D. Turnbull, and Peter B. Dervan Bioconjugate Chemistry 1994 5 (3), 242-247 DOI: 10.1021/bc00027a009
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:66919
Deposited By: Victoria Brennan
Deposited On:18 May 2016 22:33
Last Modified:26 Nov 2019 11:15

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