TY - JOUR
T1 - Crystallization and preliminary x-ray diffraction analysis of a recombinant bacterial heme oxygenase (Hmu O) from Corynebacterium diphtheriae
AU - Chu, Grace C.
AU - Park, Sam Yong
AU - Shiro, Yoshitsugu
AU - Yoshida, Tadashi
AU - Ikeda-Saito, Masao
N1 - Funding Information:
We thank Dr. Shin-ichi Adachi for his assistance in the X-ray diffraction measurements at BL44B2 (RIKEN BL2) of SPring-8 and Dr. Karen Magnus for her suggestions and the generous use of her facility. This work is supported by the National Institutes of Health Grant GM 57272 (to M.I.-S.), the SR Structural Biology Research Program in RIKEN (to Y.S.), and Grants-in-Aid for Scientific Research 09480158, 10129101, and 10044233 (to T.Y.) from the Ministry of Education, Science, Sports and Culture, Japan.
PY - 1999/6/15
Y1 - 1999/6/15
N2 - Hmu O is a 24-kDa soluble bacterial heme degradation enzyme found in the pathogen Corynebaeterium diphtheriae, the causative agent of diphtheria. Similar to the mammalian heme oxygenase, it binds hemin stoichiometrically and catalyzes the oxygen-dependent conversion of hemin to biliverdin, carbon monoxide, and free iron. Iron is an essential nutrient for bacteria and especially important for pathogenesis. Here we report the first crystallization and preliminary crystallographic study of the heme-Hmu O complex formed from hemin and a recombinant Hmu O, which was expressed in Escherichia coil from a synthetic gene based on the putative hmu O gene sequence. Crystals of the heme-Hmu O complex were obtained by the sitting drop vapor diffusion method using a precipitant solution containing 18% (w/v) PEG 8000 and 0.2 M calcium acetate in 0.1 M sodium cacodylate (pH 6.5). Using synchrotron radiation, the heme-Hmu O crystal diffracted to 2.8 Å resolution. It belongs to the monoclinic space group C2, with unit cell parameters a = 123.18 Å, b = 44.51 Å, c = 92.10 Å, and β = 123.3°. Assuming one molecule of the heme-Hmu O complex per asymmetric unit, the calculated value of V(m) is 2.89 Å3/Da.
AB - Hmu O is a 24-kDa soluble bacterial heme degradation enzyme found in the pathogen Corynebaeterium diphtheriae, the causative agent of diphtheria. Similar to the mammalian heme oxygenase, it binds hemin stoichiometrically and catalyzes the oxygen-dependent conversion of hemin to biliverdin, carbon monoxide, and free iron. Iron is an essential nutrient for bacteria and especially important for pathogenesis. Here we report the first crystallization and preliminary crystallographic study of the heme-Hmu O complex formed from hemin and a recombinant Hmu O, which was expressed in Escherichia coil from a synthetic gene based on the putative hmu O gene sequence. Crystals of the heme-Hmu O complex were obtained by the sitting drop vapor diffusion method using a precipitant solution containing 18% (w/v) PEG 8000 and 0.2 M calcium acetate in 0.1 M sodium cacodylate (pH 6.5). Using synchrotron radiation, the heme-Hmu O crystal diffracted to 2.8 Å resolution. It belongs to the monoclinic space group C2, with unit cell parameters a = 123.18 Å, b = 44.51 Å, c = 92.10 Å, and β = 123.3°. Assuming one molecule of the heme-Hmu O complex per asymmetric unit, the calculated value of V(m) is 2.89 Å3/Da.
KW - Crystallization
KW - Heme oxygenase
KW - Hmu O
KW - X-ray diffraction
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U2 - 10.1006/jsbi.1999.4122
DO - 10.1006/jsbi.1999.4122
M3 - Article
C2 - 10388628
AN - SCOPUS:0033563727
VL - 126
SP - 171
EP - 174
JO - Journal of Structural Biology
JF - Journal of Structural Biology
SN - 1047-8477
IS - 2
ER -