Properties of the detergent solubilised cytochrome c oxidase (cytochrome cbb(3)) purified from Pseudomonas stutzeri.
Cytochrome cbb(3) is a cytochrome c-oxidising isoenzyme that belongs to the superfamily of respiratory haem/copper oxidases. We have developed a purification method yielding large amounts of pure cbb(3) complex from the soil bacterium Pseudomonas stutzeri. This cytochrome cbb(3) complex consists of three subunits (ccoNOP) in a 1:1:1 stoichiometry and contains two b-type and three c-type haems. The protein complex behaves as a monomer with an overall molecular weight of 114.0+/-8.9 kDa and a s(0)(20,w) value of 8.9+/-0.3 S as determined by analytical ultracentrifugation. Crystals diffracting to 5.0 A resolution have been grown by the vapour diffusion sitting drop method to an average size of 0.1 x 0.1 x 0.3 mm. This is the first crystallisation report of a (cbb(3))-type oxidase.
Urbani
A
A
European Molecular Biology Laboratory, Structural and Computational Biology Programme, Meyerhof str. 1, D-69117 Heidelberg, Germany. a.urbani@dkfz.de
Gemeinhardt
S
S
Warne
A
A
Saraste
M
M
D000595:Amino Acid Sequence; D002458:Cell Fractionation; D003460:Crystallization; D003902:Detergents / Q000737:chemistry; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D008970:Molecular Weight; D021122:Protein Subunits; D011549:Pseudomonas / Q000737:chemistry / Q000201:enzymology*; D012995:Solubility; D014461:Ultracentrifugation; D014961:X-Ray Diffraction
D003902:Detergents; D021122:Protein Subunits; C414443:cbb3 oxidase; D003576:Electron Transport Complex IV