Abstract
Preillumination of R. rubrum membranes in the presence of 50μM phenazine methosulfate produces an inhibition of their photophosphorylating capacity.
At low pH's phosphate protects against this photo-inhibition, whereas increasing the pH eliminates this protective effect. The inhibition can be produced not only by preillumination at high pH in the presence of phenazine methosulfate, but also by preillumination at low pH (5.0) and then shifting the pH to 8.0 in the dark. We have also measured the effect of preillumination in the presence of an uncoupler such as carbonylcyanide p-trifluoromethoxyphenylhydrazone, and found that at normal pH's while photophosphorylation was protected, the uncoupler activated ATPase was inhibited pointing to a clear difference for both reactions and perhaps different structural requirements for their activities.
The purified coupling factor protein isolated from either normal or photoinactivated membrane will reconstitute normal photophosphorylation in previously uncoupled membranes but not in uncoupled membranes which were inactivated by preillumination with phenazine methosulfate prior to uncoupling.
Similar content being viewed by others
References
Avron, M.: Photophosphorylation by swiss-chard chloroplasts. Biochim. Biophys. Acta 40, 257–272 (1960)
Baccarini-Melandri, A., Fabri, E., Firstater, E., Melandri, B. A.: Energy transduction in photosynthetic bacteria. VII. Inhibition of the coupling ATPase by N-ethylmaleimide related to the energized state of the membrane. Biochim. Biophys. Acta 376, 72–81 (1975)
Binder, A., Gromet-Elhanan, Z.: Depletion and reconstitution of photophosphorylation in chromatophore membranes of Rhodospirillum rubrum. Proceeding of the 3rd International Congress on Photosynthesis (M. Avron, ed.) Vol. 2, pp. 1163–1170. Amsterdam: Elsevier Scientific Publ. Co. 1974
Boyer, P. D.: Coupling mechanisms in capture, transmission, and use of energy. Ann. Rev. Biochem. 46, 957–966 (1977)
Clayton, R. K.: Absorption spectra of photosynthetic bacteria and their chlorophylls. In: Bacterial photosynthesis (H. Gest, A. San Pietro, L. P. Vernon, eds.), pp. 495–500. Yellow Springs, Ohio: The Antioch Press 1963
Datta, D. B., Ryrie, I. J., Jagendorf, A. T.: Light-dependent modification of spinach chloroplasts coupling factor 1 by permanganate ion. J. Biol. Chem. 249, 4404–4411 (1974)
Gromet-Elhanan, Z.: Role of photophosphorylation coupling factor in energy conversion by depleted chromatophores of Rhodospirillum rubrum. J. Biol. Chem. 249, 2522–2527 (1974)
Harris, D. A., Slater, E. C.: Tightly bound nucleotides of the energy-transducing ATPase of chloroplasts and their role in photophosphorylation. Biochim. Biophys. Acta 387, 335–348 (1975)
Jagendorf, A. T.: Chloroplast membranes and coupling factor conformations. Federation Proc. 34, 1718–1722 (1975)
Kerber, N. L., Pucheu, N. L., García, A. F.: Isolation of a membrane-bound protein having coupling factor capacity as well as ADP-Pi exchange and ADPase activities from Rhodopseudomonas viridis. FEBS Lett. 72, 63–66 (1976)
Kerber, N. L., Pucheu, N. L., García, A. F.: Phenazine methosulfate mediated photoinactivation of some energy-linked reactions in Rhodospirillum rubrum. Biochem. Biophys. Res. Commun. 81, 667–671 (1978a)
Kerber, N. L., Pucheu, N. L., García, A. F.: PMS-photoinhibition in R. rubrum membranes in the presence of permeant entities affecting either the ΔΨ or ΔpH components of the proton-motive force. FEBS Lett. 94, 265–268 (1978b)
Lutz, H. U., Dahl, J. S., Bachofen, R.: Synthesis of free ATP from membrane-bound ATP in chromatophores of Rhodospirillum rubrum. Biochim. Biophys. Acta 347, 359–370 (1974)
Newton, J. W.: Preparation and solubilization of particles: chromatophores. In: Methods in enzymology (S. P. Colowick, N. O. Kaplan, eds.), Vol. V, pp. 73–77. London: Academic Press 1961
Ryrie, I. J., Jagendorf, A. T.: Inhibition of photophosphosphorylation in spinach chloroplasts by inorganic sulfate. J. Biol. Chem. 246, 582–588 (1971a)
Ryrie, I. J., Jagendorf, A. T.: An energy-linked conformational change in the coupling factor protein in chloroplasts. J. Biol. Chem. 246, 3771–3774 (1971b)
Ryrie, I. J., Jagendorf, A. T.: Correlation between a conformational change in the coupling factor protein and the high energy state in the chloroplasts. J. Biol. Chem. 247, 4453–4459 (1972)
Schuldiner, S., Padan, E., Rottemberg, H., Gromet-Elhanan, Z., Avron, M.: pH and membrane potential in bacterial chromatophores. FEBS Lett. 49, 174–177 (1974)
Slater, E. C.: Electron transfer and energy conservation. In: Dynamics of energy transducing membranes (L. Ernster, R. W. Estabrook, E. C. Slater, eds.), pp. 1–20. Amsterdam: Elsevier Scientific Publ. Co. 1974
Slooten, L., Sybesma, Ch.: Photoinactivation of Photophosphorylation and dark ATPase in Rhodospirillum rubrum chromatophores. Biochim. Biophys. Acta 449, 565–580 (1976)
Taussky, H. H., Shorr, E.: A microcolorimetric method for the determination of inorganic phosphorus. J. Biol. Chem. 202, 675–685 (1953)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pucheu, N.L., Kerber, N.L. & García, A.F. The reconstitution of photophosphorylation and the effect of inorganic phosphate at different pH's on the phenazine methosulfate photoinhibition of Rhodospirillum rubrum chromatophores. Arch. Microbiol. 120, 283–287 (1979). https://doi.org/10.1007/BF00423077
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00423077