Cytochromes of the Purple Sulfur Bacterium Ectothiorhodospira shaposhnikovii

Two c-type cytochromes (a high spin cytochrome c′ and a low spin cytochrome c-553(549) with asymmetrical α-band) and a low spin cytochrome b-558 from the purple sulfur bacterium Ectothiorhodospira shaposhnikovii were purified by ion exchange chromatography and gel filtration and characterized. Cytochrome c′ has a molecular weight of 33000 (determined by sodium dodecylsulfate electrophoresis), an isoelectric point at pH 4.5 and a redox potential of +37 mV. Absorption spectra show in the oxidized state maxima at 404 nm and in the range of 635 nm, in the reduced form maxima at 426.5 nm, 549 nm and a shoulder at 435 nm. The best purity index obtained was 0.48 (A280/A426.5). Reduced cytochrome c′ reacts with carbon monoxide. Cytochrome c-553(549) has a molecular weight of 10400, an isoelectric point at pH 5.1 and a redox potential of +248 mV. The oxidized form shows the Soret-band at 410 nm. The reduced protein reveals an asymmetrical a-band at 553 nm with a shoulder at 549 nm, the a-band at 522 nm with a shoulder at 528 nm and the γ-band at 416 nm. The best purity index obtained was 0.18 (A280/A416). Roth cytochromes could be isolated from the soluble fraction as well as from Triton X-100 treated membranes. Furthermore very low amounts of cytochromes c-553 and c-552.5 could be detected in detergent treated chromatophores. Cytochrome 6-558 - obtained from cells grown in the presence of reduced sulfur compounds in the medium - seems to be soluble or only weakly bound to the membrane. It has a molecular weight of 15800. an isoelectric point at pH 4.1 and a redox potential of -210 mV. The hemoprotein shows absorption maxima at 424.5 nm. 526.5 nm and 556.5 nm in the reduced form and at 416 nm in the oxidized state. The best purity index obtained was 0.26 (A280/A42,4.5). In addition, there were hints for the occurrence of a high spin cytochrome b′. The cytochrome pattern as well as the amount of cytochromes were dependent on growth conditions.