Isolation, crystallization and preliminary X-ray analysis of a methanol-induced corrinoid protein from Moorella thermoacetica.
A corrinoid protein was induced and overexpressed in methanol-grown cells of the thermophilic anaerobic bacterium Moorella thermoacetica. The protein was purified from cytosolic extracts. After screening for crystallization conditions and optimization, crystals were obtained that diffracted strongly on a rotating-anode X-ray source. A diffraction data set was collected and processed including reflections to 1.9 A resolution. Reflections were indexed in a primitive orthorhombic cell with unit-cell parameters a = 55.69, b = 62.74, c = 34.54 A. N-terminal amino-acid sequencing indicates that the crystals contain a C-terminal fragment of the protein.
Zhou
Weihong
W
Department of Biochemistry and Molecular Biology, The University of Georgia, Athens, Georgia 30602, USA.
Das
Amaresh
A
Habel
Jeff E
JE
Liu
Zhi-Jie
ZJ
Chang
Jessie
J
Chen
Lirong
L
Lee
Doowon
D
Nguyen
Duong
D
Chang
Shu-Huey
SH
Tempel
Wolfram
W
Rose
John P
JP
Ljungdahl
Lars G
LG
Wang
Bi-Cheng
BC
D000105:Acetyl Coenzyme A / Q000096:biosynthesis / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D045728:Corrinoids / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006094:Gram-Positive Bacteria / Q000201:enzymology* / Q000378:metabolism; D000432:Methanol / Q000378:metabolism / Q000494:pharmacology*
D001426:Bacterial Proteins; D045728:Corrinoids; D000105:Acetyl Coenzyme A; D000432:Methanol