title: Crystallization of a genetically engineered water-soluble primary penicillin target enzyme. The high molecular mass PBP2x of Streptococcus pneumoniae.
abstract: 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.
authors.affiliation: Institut de Physique, Crystallographie, Université de Liège, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Charlier
authors.forename: G
authors.initials: G
authors.lastname: Buisson
authors.forename: O
authors.initials: O
authors.lastname: Dideberg
authors.forename: J
authors.initials: J
authors.lastname: Wierenga
authors.forename: W
authors.initials: W
authors.lastname: Keck
authors.forename: G
authors.initials: G
authors.lastname: Laible
authors.forename: R
authors.initials: R
authors.lastname: Hakenbeck
mesh_terms: D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000737:chemistry*; D003460:Crystallization; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase / Q000737:chemistry*; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D011487:Protein Conformation; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D013296:Streptococcus pneumoniae / Q000201:enzymology*; D014867:Water; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D046915:Penicillin-Binding Proteins; D011994:Recombinant Proteins; D014867:Water; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase