Origin of the ATP Formed during the Light-Dependent Oxygen Uptake Catalyzed by Rhodospirillum rubrum Chromatophores

Abstract

Abstract The oxygen uptake which is observed when Rhodospirillum rubrum chromatophores are illuminated under air and in the presence of reduced 2,6-dichlorophenolindophenol (DCIP), 2,3,5,6-tetra-methyl-p-phenylenediamine (diaminodurene, DAD) or N,N′-tetramethyl-p-phenylenediamine (TMDP) depends on the electron-donor concentration according to the equation of Michaelis-Menten. The apparent Km for the donor is lowered by the electron-transfer inhibitor 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO) which causes therefore a stimulation of the rate of the reaction at non-saturating concentrations of the donors. In contrast, the ATP formation which takes place simultaneously to oxygen uptake does not show an enzyme-like dependence on donor concentration. Moreover it is inhibited by HQNO to a variable extent, depending on the particular donor present and on its concentration. Therefore it appears that the HQNO-sensitive phosphorylation is coupled to a cyclic flow which coexists and competes with the non-cyclic flow from donor to oxygen. In the presence of HQNO, substrates and uncouplers of ATP formation accelerate somewhat the rate of the oxygen uptake supported by reduced DCIP and DAD. Thus part of the HQNO-resistant phosphorylation seems to be associated with the non-cyclic flow from those two donors to oxygen. The lack of stimulation by phosphorylation or by uncoupling of the TMPD-supported oxygen uptake does not permit a conclusion as to whether this reaction is coupled to ATP forma­ tion or not. Another part of the HQNO-resistant ATP formation is independent of the presence of oxygen and appears to be associated to cyclic flows which bypass the HQNO site. This type of phosphorylation is most important in the presence of TMPD.