Preliminary molecular characterization and crystallization of mitochondrial respiratory complex II from porcine heart.
The mitochondrial respiratory complex II, or succinate:ubiquinone oxidoreductase, is an integral membrane protein complex in both the tricarboxylic acid cycle (Krebs cycle) and aerobic respiration. The gene sequences of each complex II subunit were measured by RT-PCR. N-terminal sequencing work was performed to identify the mitochondrial targeting signal peptide of each subunit. Complex II was extracted from porcine heart and purified by the ammonium sulfate precipitation method. The sample was solubilized by 0.5% (w/v) sugar detergent n-decyl-beta-D-maltoside, stabilized by 200 mM sucrose, and crystallized with 5% (w/v) poly(ethylene glycol) 4000. Important factors for the extraction, purification and crystallization of mitochondrial respiratory complex II are discussed.
Huo
Xia
X
Tsinghua-Nankai-IBP Joint Research Group for Structural Biology, Tsinghua University, Beijing 100084, China.
Su
Dan
D
Wang
Aojin
A
Zhai
Yujia
Y
Xu
Jianxing
J
Li
Xuemei
X
Bartlam
Mark
M
Sun
Fei
F
Rao
Zihe
Z
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003460:Crystallization; D017931:DNA Primers; D042963:Electron Transport Complex II / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D013552:Swine; D014158:Transcription, Genetic
D017931:DNA Primers; D042963:Electron Transport Complex II