Phenyl-azide-mediated photocrosslinking analysis of Cro-DNA interaction

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Abstract

Using phenyl-azide-mediated photocrosslinking, we show that the α carbon of amino acid 2 of the helix-turn-helix motif of bacteriophage λ Cro is within 12 Å of the bottom-strand nucleotides at positions 2 and 3 of the DNA half site in the Cro-DNA complex in solution. This result is in excellent agreement with the crystallographic structure of the Cro-DNA complex. The results of phenyl-azide-mediated photocrosslinking analysis of Cro-DNA interaction, together with the previously reported results of phenyl-azide-mediated photocrosslinking analysis of CAP-DNA interaction, establish that phenyl-azide-mediated photocrosslinking is generalizable and provide information regarding the structural requirements for phenyl-azide-mediated photocrosslinking. Comparison of the results of phenyl-azide-mediated photocrosslinking to the results of EDTA : iron-mediated affinity cleaving indicates that phenyl-azide-mediated photocrosslinking yields superior resolution.

Original languageEnglish (US)
Pages (from-to)453-460
Number of pages8
JournalJournal of molecular biology
Volume230
Issue number2
DOIs
StatePublished - Mar 20 1993

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • Azidophenacyl bromide
  • Cysteine-specific chemical modification of protein
  • Helix-turn-helix motif
  • Protein-DNA interaction

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