Studies on the Energy-coupling Sites of Photophosphorylation: V. Phosphorylation Efficiencies (P/e(2)) Associated with Aerobic Photooxidation of Artificial Electron Donors |
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Authors: | Ort D R Izawa S |
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Institution: | Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan 48824. |
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Abstract: | The rate of Hill reaction can be measured accurately as O2 uptake (the Mehler reaction) if a rapidly autoxidizable electron acceptor (e.g., methylviologen) is used. However, when an artificial electron donor-ascorbate couple (or ascorbate alone) replaces the natural donor, water, the rate of O2 consumption is no longer a reliable measure of the electron flux, because superoxide radical reactions contribute to O2 uptake. Such radical reactions, however, can be suppressed by adding enough superoxide dismutase to the reaction mixture. Indeed in all of the photosystem I- and photosystem II-donor reactions tested (except with benzidine which was tested without ascorbate added), the O2 uptake was inhibited by 30 to 50% by the addition of superoxide dismutase. The rate of phosphorylation was totally unaffected by the enzyme. The reasessment of the phosphorylation efficiencies thus made by the use of superoxide dismutase led us to the following conclusions. The phosphorylation efficiency associated with the transfer of electrons from a donor to methlylviologen (than to O2) through both photosystems II and I is practically independent of the donor used—catechol, benzidine, p-aminophenol, dicyanohydroquinone, or water. The P/e2 ratio is 1.0 ± 0.1. Only ascorbate gives a slightly lower value (P/e2 = 0.9). (NH2OH-treated, non-water-splitting chloroplasts were used for reactions with these artificial donors.) The phosphorylation efficiency associated with DCMU-insensitive, photosystem I-mediated transfer of electrons from a donor to methylviologen (then to O2) is again largely independent of the donor used, such as diaminodurene, diaminotoluene, and reduced 2,6-dichlorphenol-indophenol. The P/e2 ratio is 0.6 ± 0.08. |
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