Preparation and reconstitution of a phospholipid deficient cytochrome b6-f complex from spinach chloroplasts.
A simple, rapid procedure suitable for large scale preparation of a lipid deficient cytochrome b6-f complex from spinach chloroplasts has been developed. The procedure involves solubilization with a mixture of sodium cholate and octylglucoside, ammonium sulfate fractionation and calcium phosphate column chromatography. The purified complex contains, in nanomoles per milligram protein, 20.6 cytochrome b, 10.8 cytochrome f and 54 phospholipids. The purified complex has little plastoquinol-cytochrome c reductase activity in the absence of added lipid. Full reductase activity was reconstituted by the addition of plastoquinone prior to the addition of lipid.
D002736:Chloroplasts / Q000032:analysis*; D003573:Cytochrome b Group / Q000302:isolation & purification / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D003580:Cytochromes / Q000032:analysis; D045348:Cytochromes f; D004578:Electron Spin Resonance Spectroscopy; D008055:Lipids / Q000494:pharmacology; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000032:analysis / Q000502:physiology*; D010944:Plants / Q000032:analysis; D010971:Plastoquinone / Q000494:pharmacology; D013053:Spectrophotometry
D003573:Cytochrome b Group; D003580:Cytochromes; D008055:Lipids; D010743:Phospholipids; D045346:Cytochrome b6f Complex; D045348:Cytochromes f; D010971:Plastoquinone