Structure and function of the dihydropteroate synthase from Staphylococcus aureus

被引:150
作者
Hampele, IC [1 ]
DArcy, A [1 ]
Dale, GE [1 ]
Kostrewa, D [1 ]
Nielsen, J [1 ]
Oefner, C [1 ]
Page, MGP [1 ]
Schonfeld, HJ [1 ]
Stuber, D [1 ]
Then, RL [1 ]
机构
[1] F HOFFMANN LA ROCHE & CO LTD, DEPT PHARMA PRECLIN RES, CH-4070 BASEL, SWITZERLAND
关键词
DHPS; kinetics; sulfonamide-resistance; X-ray structure; S-aureus;
D O I
10.1006/jmbi.1997.0944
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 ape-enzyme and a binary complex with the substrate analogue hydroxymethylpterin pyrophosphate were determined at 2.2 Angstrom and 2.4 Angstrom 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. (C) 1997 Academic Press Limited.
引用
收藏
页码:21 / 30
页数:10
相关论文
共 43 条
  • [1] [Anonymous], MOL REPLACEMENT
  • [2] THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY
    BAILEY, S
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 : 760 - 763
  • [3] STRUCTURE OF CHICKEN MUSCLE TRIOSE PHOSPHATE ISOMERASE DETERMINED CRYSTALLOGRAPHICALLY AT 2.5A RESOLUTION USING AMINO-ACID SEQUENCE DATA
    BANNER, DW
    BLOOMER, AC
    PETSKO, GA
    PHILLIPS, DC
    POGSON, CI
    WILSON, IA
    CORRAN, PH
    FURTH, AJ
    MILMAN, JD
    OFFORD, RE
    PRIDDLE, JD
    WALEY, SG
    [J]. NATURE, 1975, 255 (5510) : 609 - 614
  • [4] SEQUENCE VARIATION OF THE HYDROXYMETHYLDIHYDROPTERIN PYROPHOSPHOKINASE - DIHYDROPTEROATE SYNTHASE GENE IN-LINE SO THE HUMAN MALARIA PARASITE, PLASMODIUM-FALCIPARUM, WITH DIFFERING RESISTANCE TO SULFADOXINE
    BROOKS, DR
    WANG, P
    READ, M
    WATKINS, WM
    SIMS, PFG
    HYDE, JE
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1994, 224 (02): : 397 - 405
  • [5] BRUNGER AT, 1992, XPLOR VERSION 3 1
  • [6] BURDESKA A, 1990, J MED MICROBIOL, V31, P11
  • [7] STRUCTURE OF A HYPERTHERMOPHILIC TUNGSTOPTERIN ENZYME, ALDEHYDE FERREDOXIN OXIDOREDUCTASE
    CHAN, MK
    MUKUND, S
    KLETZIN, A
    ADAMS, MWW
    REES, DC
    [J]. SCIENCE, 1995, 267 (5203) : 1463 - 1469
  • [8] Cohen Mitchell L., 1994, Trends in Microbiology, V2, P422, DOI 10.1016/0966-842X(94)90623-8
  • [9] EPIDEMIOLOGY OF DRUG-RESISTANCE - IMPLICATIONS FOR A POSTANTIMICROBIAL ERA
    COHEN, ML
    [J]. SCIENCE, 1992, 257 (5073) : 1050 - 1055
  • [10] A single amino acid substitution in Staphylococcus aureus dihydrofolate reductase determines trimethoprim resistance
    Dale, GE
    Broger, C
    DArcy, A
    Hartman, PG
    DeHoogt, R
    Jolidon, S
    Kompis, I
    Labhardt, AM
    Langen, H
    Locher, H
    Page, MGP
    Stuber, D
    Then, RL
    Wipf, B
    Oefner, C
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1997, 266 (01) : 23 - 30