Summary
Purple membrane vesicles prepared by different techniques differ widely in their morphology and ability to establish a proton gradient in the light. The procedures used to prepare active vesicles do not completely dissociate the purple membrane and thus preserve a preferential orientation of the protein, while most of the lipid is exchanged for added lipid. Responses to illumination are largely determined by the size of the vesicles and the degree to which bacteriorhodopsin is preferentially oriented. Any attempt to compare the interaction of different lipids with bacteriorhodopsin by measuring the pH response must take these factors into account.
With an improved technique we have obtained vesicles of rather uniform size and bacteriorhodopsin orientation, which accumulate protons with an initial rate of 160 ng H+ sec−1 mg−1 protein at light intensities of 106 erg cm−2 sec−1. The kinetics of the process are complex and at present insufficiently understood.
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References
Becher, B., Ebrey, T. 1976. Evidence for chromophore-chromophore (exciton) interaction in the purple membrane ofHalobacterium halobium.Biophys. J. 16:100a (Abstract TH-AM-G7)
Blaurock, A.E. 1975. Bacteriorhodopsin: A trans-membrane pump containing α-helix.J. Mol. Biol. 93:139
Blaurock, A.E., Stoeckenius, W. 1971. Structure of the purple membrane.Nature New Biol. 233:152
Bligh, E.G., Dyer, W.J. 1959. A rapid method of total lipid extraction and purification.Can. J. Biochem. 37:911
Danon, A., Stoeckenius, W. 1974. Photophosphorylation inHalobacterium halobium.Proc. Nat. Acad. Sci. USA 71:1234
Drachev, L.A., Jasaitis, A.A., Kaulen, A.D., Kondrashin, A.A., Liberman, E.A., Nemecek, I.B., Ostroumov, S.A., Semenov, A.Yu., Skulachev, V.P. 1974. Direct measurement of electric current generation by cytochrome oxidase, H+-ATPase and bacteriorhodopsin.Nature (London) 249:321
Drachev, L.A., Kaulen, A.D., Ostroumov, S.A., Skulachev, V.P. 1974. Electrogenesis by bacteriorhodopsin incorporated in a planar phospholipid membrane.FEBS Lett. 39:43
Fiske, C.H., Subbarow, Y. 1925. The colormetric determination of phosphorus.J. Biol. Chem. 66:375
Henderson, R. 1975. The structure of the purple membrane fromHalobacterium halobium: Analysis of the X-ray diffraction pattern.J. Mol. Biol. 93:123
Henderson, R., Unwin, P.N.T. 1975. Three-dimensional model of purple membrane obtained by electron microscopy.Nature (London) 257:28
Heyn, M.P., Bauer, P.-J., Dencher, N.A. 1975. A natural CD label to probe the structure of the purple membrane fromHalobacterium halobium by means of exciton coupling effects.Biochem. Biophys. Res. Commun. 67:897
Holloway, P.W. 1973. A simple procedure for removal of Triton X-100 from protein samples.Analyt. Biochem. 53:304
Hwang, S.-B., Korenbrot, J.I., Stoeckenius, W. 1977. Light-dependent proton transport by bacteriorhodopsin incorporated in an interface film.Prog. Clin. Biol. Res. (in press)
Kagawa, Y., Racker, E. 1971. Partial resolution of the enzymes catalyzing oxidative phosphorylation. XXV. Reconstitution of vesicles catalyzing32Pi-adenosine triphosphate exchange.J. Biol. Chem. 246:5477
Kates, M., Palameta, B., Joo, C.N., Kushner, D.J., Gibbons, N.E. 1966. Aliphatic diether analog of glyceride-derived lipids. IV. The occurrence of di-O-dihydrophytylglycerol ether containing lipids in extremely halophilic bacteria.Biochemistry 5:4092
Kayushin, L.P., Skulachev, V.P. 1974. Bacteriorhodopsin as an electrogenic proton pump: Reconstitution of bacteriorhodopsin proteoliposomes generating Δψ and ΔpH.FEBS Lett. 39:39
Kushwaha, S.C., Kates, M., Martin, W.G. 1975. Characterization and composition of the purple membrane and red membrane fromHalobacterium cutirubrum.Can. J. Biochem. 53:284
Kushwaha, S.C., Kates, M., Stoeckenius, W. 1976. Comparison of purple membrane fromHalobacterium cutirubrum andHalobacterium halobium.Biochim. Biophys. Acta 426:703
Lewis, A., Spoonhower, J., Bogomolni, R.A., Lozier, R.H., Stoeckenius, W. 1974. Tunable laser resonance Raman spectroscopy of bacteriorhodopsin.Proc. Nat. Acad. Sci. USA 71:4462
Lozier, R.H., Bogomolni, R.A., Stoeckenius, W. 1975. Bacteriorhodopsin: A light-driven proton pump inHalobacterium halobium.Biophys. J. 15:955
Lozier, R.H., Niederberger, W., Bogomolni, R.A., Hwang, S.-B., Stoeckenius, W. 1976. Kinetics and stoichiometry of light-induced proton release and uptake from purple membrane fragments,Halobacterium halobium cell envelopes, and phospholipid vesicles containing oriented purple membrane.Biochim. Biophys. Acta 440:545
Mitchell, P. 1969. Oriented chemical reactions and ion movements in membranes.In: The Molecular Basis of Membrane Functions. D.C. Tosteson editor. p. 483. Prentice Hall Inc., Englewood Cliffs, N.J.
Miyamoto, V.K., Stoeckenius, W. 1971. Preparation and characteristics of lipid vesicles.J. Membrane Biol. 4:252
Moor, H., Mühlethaler, K. 1963. Fine structure in frozen-etched cells.J. Cell Biol. 17:609
Oesterhelt, D., Stoeckenius, W. 1971. Rhodopsin-like protein from the purple membrane ofHalobacterium halobium.Nature New Biol. 233:149
Oesterhelt, D., Stoeckenius, W. 1973. Function of a new photoreceptor membrane.Proc. Nat. Acad. Sci. USA 70:2853
Oesterhelt, D., Stoeckenius, W. 1974. Isolation of the cell membrane ofHalobacterium halobium and its fractionation into red and purple membrane.In: Methods in Enzymology, Vol. 31, Biomembranes, Part A. S. Fleischer and R. Estabrook, editors. Academic Press, New York
Racker, E. 1973. A new procedure for the reconstitution of biological active phospholipid vesicles.Biochem. Biophys. Res. Commun. 55:224
Racker, E., Hinkle, P.C. 1974. Effect of temperature on the function of a proton pump.J. Membrane Biol. 17:181
Racker, E., Stoeckenius, W. 1974. Reconstitution of purple membrane vesicles catalyzing light-driven proton uptake and adenosine triphosphate formation.J. Biol. Chem. 249:662
Shipley, R.A., Clark, R.E. 1972. Tracer Methods for In vivo Kinetics. Academic Press, New York & London
Stoeckenius, W. 1976. Structure of biological membranes: Bacteriorhodopsin and the purple membrane. Proceedings of a Conference held in Rome, April 14–19, 1975; Pontifical Academy of Sciences “Biological and Artificial Membranes and Desalination of Water”
Stoeckenius, W., Hwang, S.-B., Korenbrot, J. 1977. Proton translocation by bacteriorhodopsin in model systems. Nobel Symposium 34: The Structure of Biological Membranes, S. Abrahamson and I. Pescher, editors. pp. 479–496. Plenum Press, New York
Stoeckenius, W., Rowen, R. 1967. A morphological study ofHalobacterium halobium and its lysis in media of low salt concentration.J. Cell Biol. 34:365
Unwin, P.N.T., Henderson, R. 1975. Molecular structure determination by electron microscopy of unstained crystalline specimens.J. Mol. Biol. 94:425
Yaguzhinsky, L.S., Boguslavsky, L.I., Volkov, A.G., Rakhmaninova, A.B. 1976. Synthesis of ATP coupled with action of membrane protonic pumps at the octane-water interfaces.Nature (London) 259:494
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Hwang, SB., Stoeckenius, W. Purple membrane vesicles: Morphology and proton translocation. J. Membrain Biol. 33, 325–350 (1977). https://doi.org/10.1007/BF01869523
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DOI: https://doi.org/10.1007/BF01869523