Crystallization of a genetically engineered water-soluble primary penicillin target enzyme. The high molecular mass PBP2x of Streptococcus pneumoniae

J Mol Biol. 1993 Aug 5;232(3):1007-9. doi: 10.1006/jmbi.1993.1452.


A genetically engineered water-soluble derivative of PBP2x of Streptococcus pneumoniae has been produced, purified and crystallized in a form suitable for X-ray diffraction analysis. The best crystals have been grown at 15 degrees C, from solutions containing 8% polyethylene glycol 10,000 at pH values ranging from 3.9 to 6.0. These crystals diffract to a resolution of 3.5 A and have a space group P6(1)22 (or enantiomorph) with unit cell dimensions of a = b = 162.2 A, c = 171.8 A, alpha = beta = 90 degrees, gamma = 120 degrees. The molecular mass and cell dimensions suggest that there is one molecule of enzyme per asymmetric unit. The breakdown of a chromogenic cephalosporin derivative diffused into a crystal reveals clearly that the enzyme is active in the crystalline state.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins*
  • Carrier Proteins / chemistry*
  • Crystallization
  • Hexosyltransferases*
  • Muramoylpentapeptide Carboxypeptidase / chemistry*
  • Penicillin-Binding Proteins
  • Peptidyl Transferases*
  • Protein Conformation
  • Protein Engineering
  • Recombinant Proteins / chemistry
  • Solubility
  • Streptococcus pneumoniae / enzymology*
  • Water
  • X-Ray Diffraction


  • Bacterial Proteins
  • Carrier Proteins
  • Penicillin-Binding Proteins
  • Recombinant Proteins
  • Water
  • Peptidyl Transferases
  • Hexosyltransferases
  • Muramoylpentapeptide Carboxypeptidase