Properties of cytochrome bd plastoquinol oxidase from the cyanobacterium Synechocystis sp. PCC 6803.
In the aerobic respiratory chain of the cyanobacterium Synechocystis sp. PCC 6803, cytochrome c oxidase serves as a major terminal oxidase while cyanide-resistant cytochrome bd serves as an alternative oxidase and evades the over-reduction of the plastoquinone pool under stress conditions. Here we expressed Synechocystis cytochrome bd in Escherichia coli and characterized enzymatic and spectroscopic properties. Cyanobacterial cytochrome bd showed the higher activity with ubiquinols than with decyl-plastoquinol and K(m) values for quinols were 2-fold smaller than those of E. coli cytochrome bd (CydAB). The dioxygen reduction site was resistant to cyanide as in E. coli oxidase while the quinol oxidation site was more sensitive to antimycin A and quinolone inhibitors. Spectroscopic analysis showed the presence of the haem b(595)-d binuclear centre but the sequence analysis indicates that cyanobacterial cytochrome bd is structurally related to cyanide-insensitive oxidase (CioAB), which does not show typical spectral changes upon reduction and ligand binding. Our data indicate that cyanobacterial cytochrome bd has unique enzymatic and structural properties and we hope that our findings will help our understanding the role and properties of CydAB and CioAB quinol oxidases in other bacterial species.
Mogi
Tatsushi
T
Department of Biomedical Chemistry, Graduate School of Medicine, the University of Tokyo, Bunkyo-ku, Tokyo, Japan. tmogi@m.u-tokyo.ac.jp
Miyoshi
Hideto
H
D000595:Amino Acid Sequence; D003580:Cytochromes / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction / Q000187:drug effects; D010100:Oxygen / Q000378:metabolism; D010802:Phylogeny; D010971:Plastoquinone / Q000031:analogs & derivatives* / Q000378:metabolism; D011808:Quinone Reductases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D013057:Spectrum Analysis; D046939:Synechocystis / Q000187:drug effects / Q000201:enzymology*
D003580:Cytochromes; D004791:Enzyme Inhibitors; C003165:plastoquinol; D011808:Quinone Reductases; C030914:ubiquinol-1 oxidase; D010971:Plastoquinone; D010100:Oxygen