Crystallization and preliminary X-ray diffraction analysis of fatty-acid hydroxylase cytochrome P450BSbeta from Bacillus subtilis.
Cytochrome P450 isolated from Bacillus subtilis (P450BSbeta; MW 48 kDa) catalyzes the hydroxylation of long-chain fatty acids at the alpha and beta positions using H(2)O(2) as an oxidant. Crystals of the substrate-free form of P450BSbeta belonging to the trigonal space group P3(2)21 or P3(1)21 were obtained by the sitting-drop vapour-diffusion method using a precipitate solution consisting of 10%(w/v) PEG 4000 and 50 mM MES pH 6.8. Another crystal form, belonging to the rhombohedral space group R3 or R32, was obtained from precipitate solution consisting of 10% PEG 4000, 0.15 mM magnesium acetate and 50 mM MES pH 6.5 in the presence of 2 mM myristic acid (substrate). Using synchrotron radiation, both P450BSbeta crystals diffracted to 2.5 A resolution. Bijvoet and dispersive anomalous difference Patterson maps show a clear peak corresponding to the haem iron.
Lee
Dong Sun
DS
RIKEN Harima Institute/Spring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo, Hyogo 679-5148, Japan.
Yamada
Akari
A
Matsunaga
Isamu
I
Ichihara
Kosuke
K
Adachi
Shin-ichi
S
Park
Sam-Yong
SY
Shiro
Yoshitsugu
Y
D001412:Bacillus subtilis / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D010544:Peroxidases / Q000737:chemistry* / Q000378:metabolism
D005227:Fatty Acids; D003577:Cytochrome P-450 Enzyme System; D010544:Peroxidases; C437763:cytochrome P-450 CYP152A1 (Bacillus subtilis)