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An electrical probe of protein-DNA interactions on DNA-modified surfaces

Boon, Elizabeth M. and Salas, Julia E. and Barton, Jacqueline K. (2002) An electrical probe of protein-DNA interactions on DNA-modified surfaces. Nature Biotechnology, 20 (3). pp. 282-286. ISSN 1087-0156. doi:10.1038/nbt0302-282. https://resolver.caltech.edu/CaltechAUTHORS:20150401-111610690

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Abstract

DNA charge transport chemistry is found to provide a sensitive method for probing protein-dependent changes in DNA structure and enzymatic reactions. Here we describe the development of an electrochemical assay of protein binding to DNA-modified electrodes based upon the detection of associated perturbations in DNA base stacking. Gold electrode surfaces that were modified with loosely packed DNA duplexes, covalently crosslinked to a redox-active intercalator and containing the binding site of the test protein, were constructed. Charge transport through DNA as a function of protein binding was then assayed. Substantial attenuation in current is seen in the presence of the base-flipping enzymes HhaI methylase and uracil DNA glycosylase, as well as with TATA-binding protein. When restriction endonuclease PvuII (R.PvuII) binds to its methylated target, little base-stacking perturbation occurs and little diminution in current flow is observed. Importantly, the kinetics of restriction by R.PvuII of its nonmethylated target is also easily monitored electrochemically. This approach should be generally applicable to assaying protein–DNA interactions and reactions on surfaces.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1038/nbt0302-282 DOIArticle
http://www.nature.com/nbt/journal/v20/n3/full/nbt0302-282.htmlPublisherArticle
http://rdcu.be/csa6PublisherFree ReadCube access
ORCID:
AuthorORCID
Boon, Elizabeth M.0000-0003-1891-839X
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2002 Nature Publishing Group. We are grateful to the NIH for their financial support (GM61077) and for an NRSA predoctoral fellowship (E.M.B.). We thank also the Caltech SURF program for a summer undergraduate fellowship (J.E.S.) and Donato Ceres for assistance in obtaining the scanning force microscopy images.
Funders:
Funding AgencyGrant Number
NIHGM61077
NIH Predoctoral FellowshipUNSPECIFIED
Caltech Summer Undergraduate Research Fellowship (SURF)UNSPECIFIED
Issue or Number:3
DOI:10.1038/nbt0302-282
Record Number:CaltechAUTHORS:20150401-111610690
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150401-111610690
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:56284
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Apr 2015 19:05
Last Modified:10 Nov 2021 20:57

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