TY - JOUR
T1 - Purification, characterization and crystallization of menaquinol:fumarate oxidoreductase from the green filamentous photosynthetic bacterium Chloroflexus aurantiacus
AU - Xin, Yueyong
AU - Lu, Yih Kuang
AU - Fromme, Raimund
AU - Fromme, Petra
AU - Blankenship, Robert E.
N1 - Funding Information:
We thank Drs. Daniel Brune and John Lopez for their technical support for the MS measurement, Dr. Russ Lobrutto for EPR measurement, Dr. Meitian Wang for crystal screening and the staff of BL 8.2.1 in ALS and ID 19 in APS for X-ray diffraction measurements. This work was supported by grants to REB from the Exobiology Program from NASA and from the Molecular Biochemistry Program from NSF (Grant 0646621).
PY - 2009/2
Y1 - 2009/2
N2 - The integral membrane protein complex, menaquinol:fumarate oxidoreductase (mQFR) has been purified, identified and characterized from the thermophilic green filamentous anoxygenic photosynthetic bacterium Chloroflexus aurantiacus. The complex is composed of three subunits: a 74 kDa flavoprotein that contains a covalently bound flavin adenine dinucleotide, a 28 kDa iron-sulfur cluster-containing polypeptide, and a 27 kDa transmembrane polypeptide, which is also the binding site of two b-type hemes and two menaquinones. The purified complex has an apparent molecular mass of 260 kDa by blue-native PAGE, which is indicative of a native homodimeric form. The isolated complex is active in vitro in both fumarate reduction and succinate oxidation. It has been analyzed by visible absorption, redox titration, chemical analysis and EPR spectroscopy. In addition, phylogenetic analysis shows that the QFR of both C. aurantiacus and Chlorobium tepidum are most closely related to those found in the delta-proteobacteria. The purified enzyme was crystallized and X-ray diffraction data obtained up to 3.2 Å resolution.
AB - The integral membrane protein complex, menaquinol:fumarate oxidoreductase (mQFR) has been purified, identified and characterized from the thermophilic green filamentous anoxygenic photosynthetic bacterium Chloroflexus aurantiacus. The complex is composed of three subunits: a 74 kDa flavoprotein that contains a covalently bound flavin adenine dinucleotide, a 28 kDa iron-sulfur cluster-containing polypeptide, and a 27 kDa transmembrane polypeptide, which is also the binding site of two b-type hemes and two menaquinones. The purified complex has an apparent molecular mass of 260 kDa by blue-native PAGE, which is indicative of a native homodimeric form. The isolated complex is active in vitro in both fumarate reduction and succinate oxidation. It has been analyzed by visible absorption, redox titration, chemical analysis and EPR spectroscopy. In addition, phylogenetic analysis shows that the QFR of both C. aurantiacus and Chlorobium tepidum are most closely related to those found in the delta-proteobacteria. The purified enzyme was crystallized and X-ray diffraction data obtained up to 3.2 Å resolution.
KW - Characterization
KW - Chloroflexus aurantiacus
KW - Crystallization
KW - Electron transfer chain
KW - Menaquinol:fumarate oxidoreductase
KW - Purification
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U2 - 10.1016/j.bbabio.2008.11.010
DO - 10.1016/j.bbabio.2008.11.010
M3 - Article
C2 - 19103151
AN - SCOPUS:58649090657
SN - 0005-2728
VL - 1787
SP - 86
EP - 96
JO - Biochimica et Biophysica Acta - Bioenergetics
JF - Biochimica et Biophysica Acta - Bioenergetics
IS - 2
ER -