Minimization of a Protein−DNA Dimerizer
Abstract
A protein−DNA dimerizer constructed from a DNA-binding polyamide and the peptide FYPWMKG facilitates the binding of a natural transcription factor Exd to an adjacent DNA site. The Exd binding domain can be reduced to a dipeptide WM attached to the polyamide through an ε-aminohexanoic acid linker with retention of protein−DNA dimerizer activity. Screening a library of analogues indicated that the tryptophan indole moiety is more important than methionine's side chain or the N-terminal acetamide. Remarkably, switching the stereochemistry of the tryptophan residue (l to d) stabilizes the dimerizer•Exd•DNA ternary complex at 37 °C. These observations provide design principles for artificial transcription factors that may function in concert with the cellular regulatory circuitry.
Additional Information
© 2007 American Chemical Society. Received November 7, 2006. Published on Web 02/10/2007. This work was supported by the National Institute of Health (Research Grants to P.B.D. and A.Z.A). A DAAD postdoctoral fellowship (to H.D.A) is gratefully acknowledged. We thank Professor A. L. Aggarwal (Mount Sinai School of Medicine, New York) for a donation of the plasmid encoding the Exd DNA binding domain and Drs. P. Snow and G. Hathaway (Beckman Institute, California Institute of Technology) for technical assistance with protein expression, purification, and identification.Attached Files
Accepted Version - nihms-277612.pdf
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Additional details
- PMCID
- PMC3064071
- Eprint ID
- 66751
- Resolver ID
- CaltechAUTHORS:20160509-110447840
- NIH
- Deutscher Akademischer Austauschdienst (DAAD)
- Created
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2016-05-17Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field