Structure and function of the dihydropteroate synthase from Staphylococcus aureus.

The gene encoding the dihydropteroate synthase of staphylococcus aureus has been cloned, sequenced and expressed in Escherichia coli. The protein has been purified for biochemical characterization and X-ray crystallographic studies. The enzyme is a dimer in solution, has a steady state kinetic mechanism that suggests random binding of the two substrates and half-site reactivity. The crystal structure of apo-enzyme and a binary complex with the substrate analogue hydroxymethylpterin pyrophosphate were determined at 2.2 A and 2.4 A resolution, respectively. The enzyme belongs to the group of "TIM-barrel" proteins and crystallizes as a non-crystallographic dimer. Only one molecule of the substrate analogue bound per dimer in the crystal. Sequencing of nine sulfonamide-resistant clinical isolates has shown that as many as 14 residues could be involved in resistance development. The residues are distributed over the surface of the protein, which defies a simple interpretation of their roles in resistance. Nevertheless, the three-dimensional structure of the substrate analogue binary complex could give important insight into the molecular mechanism of this enzyme.

[1]  W. Dallas,et al.  Cloning, sequencing, and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100 , 1992, Journal of bacteriology.

[2]  D. Rees,et al.  Structure of a hyperthermophilic tungstopterin enzyme, aldehyde ferredoxin oxidoreductase , 1995, Science.

[3]  J. E. Hyde,et al.  Sequence variation of the hydroxymethyldihydropterin pyrophosphokinase: dihydropteroate synthase gene in lines of the human malaria parasite, Plasmodium falciparum, with differing resistance to sulfadoxine. , 1994, European journal of biochemistry.

[4]  Mitchell L. Cohen,et al.  Antimicrobial resistance: prognosis for public health. , 1994, Trends in microbiology.

[5]  H. Nikaido,et al.  Prevention of drug access to bacterial targets: permeability barriers and active efflux. , 1994, Science.

[6]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[7]  G. Hitchings,et al.  Inhibition of folate biosynthesis and function as a basis for chemotherapy. , 1965, Advances in enzymology and related areas of molecular biology.

[8]  D. Stüber,et al.  System for High-Level Production in Escherichia coli and Rapid Purification of Recombinant Proteins: Application to Epitope Mapping, Preparation of Antibodies, and Structure—Function Analysis , 1990 .

[9]  G. Petsko,et al.  Structure of chicken muscle triose phosphate isomerase determined crystallographically at 2.5Å resolution: using amino acid sequence data , 1975, Nature.

[10]  Axel T. Brunger,et al.  Model bias in macromolecular crystal structures , 1992 .

[11]  A. Jenkins,et al.  Transformational exchanges in the dihydropteroate synthase gene of Neisseria meningitidis: a novel mechanism for acquisition of sulfonamide resistance , 1992, Journal of bacteriology.

[12]  T. Prangé,et al.  On the Preparation and X-Ray Data-Collection of Isomorphous Xenon Derivatives , 1994 .

[13]  M. Page,et al.  A single amino acid substitution in Staphylococcus aureus dihydrofolate reductase determines trimethoprim resistance. , 1997, Journal of molecular biology.

[14]  I. Kohl,et al.  Frequency and transferability of trimethoprim and sulfonamide resistance in methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. , 1992, Journal of chemotherapy.

[15]  R. Huber,et al.  Accurate Bond and Angle Parameters for X-ray Protein Structure Refinement , 1991 .

[16]  S. Lacks,et al.  Sulfonamide resistance in Streptococcus pneumoniae: DNA sequence of the gene encoding dihydropteroate synthase and characterization of the enzyme , 1987, Journal of bacteriology.

[17]  J. Navaza,et al.  AMoRe: an automated package for molecular replacement , 1994 .

[18]  Wolfgang Kabsch,et al.  Evaluation of Single-Crystal X-ray Diffraction Data from a Position-Sensitive Detector , 1988 .

[19]  Sung-Hou Kim,et al.  Sparse matrix sampling: a screening method for crystallization of proteins , 1991 .

[20]  O. Sköld,et al.  Sulfonamide resistance in Neisseria meningitidis as defined by site-directed mutagenesis could have its origin in other species , 1995, Journal of bacteriology.

[21]  A. Peregrine,et al.  Epidemiology of drug resistance , 1992 .

[22]  P. Rådström,et al.  RSF1010 and a conjugative plasmid contain sulII, one of two known genes for plasmid-borne sulfonamide resistance dihydropteroate synthase , 1988, Antimicrobial Agents and Chemotherapy.

[23]  D. Stammers,et al.  The multifunctional folic acid synthesis fas gene of Pneumocystis carinii appears to encode dihydropteroate synthase and hydroxymethyldihydropterin pyrophosphokinase. , 1992, Gene.

[24]  H. Neu,et al.  The Crisis in Antibiotic Resistance , 1992, Science.

[25]  R. Ferone,et al.  The characteristics and significance of sulfonamides as substrates for Escherichia coli dihydropteroate synthase. , 1979, The Journal of biological chemistry.

[26]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[27]  J Davies,et al.  Inactivation of antibiotics and the dissemination of resistance genes. , 1994, Science.

[28]  Thomas J. White,et al.  PCR protocols: a guide to methods and applications. , 1990 .

[29]  Mitchell L. Cohen.  Epidemiology of Drug Resistance: Implications for a Post—Antimicrobial Era , 1992, Science.

[30]  I. Crawford,et al.  An apparent Bacillus subtilis folic acid biosynthetic operon containing pab, an amphibolic trpG gene, a third gene required for synthesis of para-aminobenzoic acid, and the dihydropteroate synthase gene , 1990, Journal of bacteriology.

[31]  Paul R. Gerber,et al.  Peptide mechanics: A force field for peptides and proteins working with entire residues as smallest units , 1992 .

[32]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.