Minimization of a protein-DNA dimerizer

J Am Chem Soc. 2007 Mar 7;129(9):2660-8. doi: 10.1021/ja067971k. Epub 2007 Feb 10.

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 epsilon-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 degrees C. These observations provide design principles for artificial transcription factors that may function in concert with the cellular regulatory circuitry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetamides / chemistry
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • DNA / chemistry*
  • DNA / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism
  • Dimerization
  • Dipeptides / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Indoles / chemistry
  • Molecular Conformation
  • Molecular Sequence Data
  • Nylons / chemistry
  • Proteins / chemistry*
  • Proteins / metabolism
  • Temperature
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism
  • Tryptophan / chemistry

Substances

  • Acetamides
  • DNA-Binding Proteins
  • Dipeptides
  • Indoles
  • Nylons
  • Proteins
  • Transcription Factors
  • Tryptophan
  • acetamide
  • DNA