CaltechAUTHORS
  A Caltech Library Service

Protein stitchery: Design of a protein for selective binding to a specific DNA sequence

Park, Changmoon and Campbell, Judith L. and Goddard, William A., III (1992) Protein stitchery: Design of a protein for selective binding to a specific DNA sequence. Proceedings of the National Academy of Sciences of the United States of America, 89 (19). pp. 9094-9096. ISSN 0027-8424. PMCID PMC50071. doi:10.1073/pnas.89.19.9094. https://resolver.caltech.edu/CaltechAUTHORS:20141216-142043196

[img]
Preview
PDF - Published Version
See Usage Policy.

890kB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20141216-142043196

Abstract

We present a general strategy for designing proteins to recognize DNA sequences and illustrate this with an example based on the "Y-shaped scissors grip" model for leucine-zipper gene-regulatory proteins. The designed protein is formed from two copies, in tandem, of the basic (DNA binding) region of v-Jun. These copies are coupled through a tripeptide to yield a "dimer" expected to recognize the sequence TCATCGATGA (the v-Jun-v-Jun homodimer recognizes ATGACTCAT). We synthesized the protein and oligonucleotides containing the proposed binding sites and used gel-retardation assays and DNase I footprinting to establish that the dimer binds specifically to the DNA sequence TCATCGATGA but does not bind to the wild-type DNA sequences, nor to oligonucleotides in which the recognition half-site is modified by single-base changes. These results also provide strong support for the Y-shaped scissors grip model for binding of leucine-zipper proteins.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.89.19.9094DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC50071/PublisherArticle
ORCID:
AuthorORCID
Goddard, William A., III0000-0003-0097-5716
Additional Information:© 1992 National Academy of Sciences. Contributed by William A. Goddard III, June 3, 1992. We thank Prof. Mel Simon (California Institute of Technology) for helpful suggestions and discussion and for use of his laboratory resources (funded by National Science Foundation Grant-DMB-90-18536). We also thank Prof. David Eisenberg (University of California, Los Angeles) for helpful comments. This research was supported by a grant from Department of Energy-Advanced Industrial Concepts Division and by funding from the Materials and Molecular Simulation Center of the Beckman Institute. This is contribution number 8547 from the Division of Chemistry and Chemical Engineering.
Funders:
Funding AgencyGrant Number
NSFDMB-90-18536
Department of Energy (DOE)UNSPECIFIED
Caltech Beckman InstituteUNSPECIFIED
Other Numbering System:
Other Numbering System NameOther Numbering System ID
Caltech Division of Chemistry and Chemical Engineering8547
Issue or Number:19
PubMed Central ID:PMC50071
DOI:10.1073/pnas.89.19.9094
Record Number:CaltechAUTHORS:20141216-142043196
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20141216-142043196
Official Citation:Protein stitchery: design of a protein for selective binding to a specific DNA sequence. C Park, J L Campbell, and W A Goddard, 3rd PNAS 1992 89 (19) 9094-9096; doi:10.1073/pnas.89.19.9094
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52891
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:16 Dec 2014 22:39
Last Modified:10 Nov 2021 19:45

Repository Staff Only: item control page