Crystallization of ccdB

Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):975-81. doi: 10.1107/s0907444997011554.

Abstract

CcdB is a small dimeric protein that poisons DNA-topoisomerase II complexes. Its crystallization properties in terms of precipitant type, precipitant concentration, pH and protein concentration have been investigated leading to a novel crystal form which, in contrast to previously reported crystals, is suitable for structure determination using the multiple isomorphous replacement (MIR) method. The space group of this new form is C2, with unit-cell parameters a = 74.94, b = 36.24, c = 35.77 A, beta = 115.27 degrees. The asymmetric unit contains a single monomer. Flash-frozen crystals diffract to at least 1.5 A resolution, while room-temperature diffraction can be observed up to 1.6 A. The double mutant S74C/G77Q, which acts as a super-killer, crystallizes in space group I222 (or I212121) with unit-cell dimensions a = 105.58, b = 105.80, c = 91.90 A. These crystals diffract to 2.5 A resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification
  • Bacterial Toxins / chemistry*
  • Bacterial Toxins / genetics
  • Bacterial Toxins / isolation & purification
  • Crystallization
  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / isolation & purification
  • Escherichia coli
  • Mutagenesis, Site-Directed
  • Protein Conformation*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification*
  • Topoisomerase II Inhibitors

Substances

  • Bacterial Proteins
  • Bacterial Toxins
  • CcdB protein, Plasmid F
  • Enzyme Inhibitors
  • Recombinant Fusion Proteins
  • Topoisomerase II Inhibitors