Crystallization and preliminary X-ray diffraction analysis of a cytochrome P450 (CYP119) from Sulfolobus solfataricus

被引:27
作者
Park, SY
Yamane, K
Adachi, S
Shiro, Y
Weiss, KE
Sligar, SG
机构
[1] RIKEN, Harima Inst, Sayo, Hyogo 6795148, Japan
[2] Univ Illinois, Beckman Inst Adv Sci & Technol, Urbana, IL 61801 USA
来源
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2000年 / 56卷
关键词
D O I
10.1107/S0907444900008234
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
CYP119 is a cytochrome P450 with a molecular weight of 43 kDa which has been isolated from the thermophilic archaeon Sulfolobus solfataricus. This enzyme is extremely stable to high temperature and high pressure. The first crystallization and preliminary crystallographic study of CYP119 is reported here. Crystals of CYP119 were obtained by the sitting-drop vapour-diffusion method using a precipitant solution containing 20%(w/v) PEG 4000 and 0.2 M sodium thiocyanate at pH 6.4. Using synchrotron radiation, the CYP119 crystal diffracted to 1.84 Angstrom resolution. It belongs to the tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 86.17 (0.07), c = 221.11 (0.04) Angstrom, in which the numbers in parentheses describe the standard deviations. Assuming two molecules of the CYP119 per asymmetric unit, the calculated molar volume (V-m) is 2.38 Angstrom(3) Da(-1). Bijvoet and dispersive anomalous difference Patterson maps show a clear peak corresponding to the haem irons. The complete crystallographically defined structure is currently in progress using MIR (multiple isomorphous replacement) and MAD (multiwavelength anomalous diffraction) techniques.
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收藏
页码:1173 / 1175
页数:3
相关论文
共 9 条
  • [1] ADACHI S, 1996, SPRING 8 ANN REPORT, P239
  • [2] THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY
    BAILEY, S
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 : 760 - 763
  • [3] LESLIE AGW, 1994, MOSFLM USERS GUIDE
  • [4] SOLVENT CONTENT OF PROTEIN CRYSTALS
    MATTHEWS, BW
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1968, 33 (02) : 491 - +
  • [5] Characterization of a cytochrome P450 from the acidothermophilic archaea Sulfolobus solfataricus
    McLean, MA
    Maves, SA
    Weiss, KE
    Krepich, S
    Sligar, SG
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 252 (01) : 166 - 172
  • [6] Crystal structure of nitric oxide reductase from denitrifying fungus Fusarium oxysporum
    Park, SY
    Shimizu, H
    Adachi, S
    Nakagawa, A
    Tanaka, I
    Nakahara, K
    Shoun, H
    Obayashi, E
    Nakamura, H
    Iizuka, T
    Shiro, Y
    [J]. NATURE STRUCTURAL BIOLOGY, 1997, 4 (10) : 827 - 832
  • [7] Peterson Julian A., 1995, P151
  • [8] Poulos Thomas L., 1995, P125
  • [9] Cloning of a potential cytochrome P450 from the archaeon Sulfolobus solfataricus
    Wright, RL
    Harris, K
    Solow, B
    White, RH
    Kennelly, PJ
    [J]. FEBS LETTERS, 1996, 384 (03): : 235 - 239