2OVG : Lambda Cro Q27P/A29S/K32Q triple mutant at 1.35 A in space group P3221

  • Branwen M. Hall (Contributor)
  • Sue A. Roberts (Contributor)
  • Annie Heroux (Contributor)
  • Matthew Hj Cordes (Contributor)



Experimental Technique/Method:X-RAY DIFFRACTION
Release Date:2008-01-08
Deposition Date:2007-02-13
Revision Date:2011-07-13#2017-10-18
Molecular Weight:7693.79
Macromolecule Type:Protein
Residue Count:66
Atom Site Count:514

Previously reported crystal structures of free and DNA-bound dimers of lambda Cro differ strongly (about 4 A backbone rmsd), suggesting both flexibility of the dimer interface and induced-fit protein structure changes caused by sequence-specific DNA binding. Here, we present two crystal structures, in space groups P3(2)21 and C2 at 1.35 and 1.40 A resolution, respectively, of a variant of lambda Cro with three mutations in its recognition helix (Q27P/A29S/K32Q, or PSQ for short). One dimer structure (P3(2)21; PSQ form 1) resembles the DNA-bound wild-type Cro dimer (1.0 A backbone rmsd), while the other (C2; PSQ form 2) resembles neither unbound (3.6 A) nor bound (2.4 A) wild-type Cro. Both PSQ form 2 and unbound wild-type dimer crystals have a similar interdimer beta-sheet interaction between the beta1 strands at the edges of the dimer. In the former, an infinite, open beta-structure along one crystal axis results, while in the latter, a closed tetrameric barrel is formed. Neither the DNA-bound wild-type structure nor PSQ form 1 contains these interdimer interactions. We propose that beta-sheet superstructures resulting from crystal contact interactions distort Cro dimers from their preferred solution conformation, which actually resembles the DNA-bound structure. These results highlight the remarkable flexibility of lambda Cro but also suggest that sequence-specific DNA binding may not induce large changes in the protein structure.
Date made available2008

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