On the question of interheme electron transfer in the chloroplast cytochrome b6 in situ.
The redox properties, the site of action of the inhibitor NQNO, and the question of interheme transfer in the chloroplast cytochrome b6 have been examined with regard to the role of the b6-f complex in quinol oxidation and H+ translocation. (i) The two hemes of the cytochrome ba and bp, have similar (delta Em less than or equal to 50 mV) oxidation-reduction midpoint potentials that are pH-independent in the range pH 6.5-8.0 (Em7 = -40 mV) but are pH dependent below this range with an estimated pK = 6.7. (ii) Only half of cytochrome b6, the stromal-side heme, ba, was reducible by NADPH and ferredoxin. (iii) The 2-3-fold increase (to 0.60 +/- 0.09 heme/600 Chl) in the amplitude of flash-induced cytochrome reduction caused by NQNO was not affected when heme ba was initially reduced, implying that NQNO affects flash reduction at the site of heme bp. (iv) Multiple light flashes did not increase the amplitude of b6 reduction in the presence or absence of NQNO or show binary oscillations. Together with localization of a site of action of NQNO near heme bp, these data provide no evidence for efficient electron transfer from heme bp to heme ba as specified by the Q cycle model. (v) NQNO interaction with heme bp does not block its oxidation, since reoxidation of the flash-reduced cytochrome in its presence or absence was 4-5 times faster (t1/2 approximately 30 ms) when heme ba was reduced. The faster oxidation of the photoreduced cytochrome after NADPH-Fd reduction of heme ba indicates that the oxidation of ba and bp may be cooperative.
Furbacher
P N
PN
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Girvin
M E
ME
Cramer
W A
WA
D000968:Antimycin A / Q000494:pharmacology; D002736:Chloroplasts / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D004579:Electron Transport; D006418:Heme / Q000502:physiology; D006912:Hydroxyquinolines / Q000494:pharmacology; D010084:Oxidation-Reduction; D010766:Phosphorylation; D010944:Plants
D003573:Cytochrome b Group; D006912:Hydroxyquinolines; C001333:2-(n-heptyl)-4-hydroxyquinoline N-oxide; D006418:Heme; D000968:Antimycin A; D045346:Cytochrome b6f Complex