Summary
We have studied isolated purple membrane from Halobacterium halobium strain S9 and white membrane from the mutant strain JW-5, making careful measurements of the proton to M412 stoichiometry of our preparations, and also examining them by Quasi Elastic Light Scattering (QELS) and by negative staining electron microscopy (EM), to assess extent of aggregation and hydrodynamic radius.
In the past decade there have been varying reports of the stoichiometry of proton release from purple membrane in suspension and protons pumped after its incorporation into phospholipid vesicles. Interpretation of the results, especially when by a group other than that which carried out the experiments, has sometimes been clouded by a misunderstanding of the differences between the various measurements involved and also by a lack of recognition that the aggregation state of membranes will affect the measured ratios.
In an attempt to resolve these misunderstandings, we have investigated the H+/M412 ratio, measured in suspensions of purple membranes by the laser flash induced response of pH indicator dyes. We have found this ratio to reflect the aggregation state of the membrane, assessed on the same preparations by QELS (and EM, see accompanying chapter: Lefort-Tran et al.). We have also found our measurements of proton release stoichiometry on retinal reconstituted white membrane suspensions to give much higher H+/M412 ratios than the same measurements on purple membrane suspensions.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Blaurock, A.E., Stoeckenius, W., Oesterhelt, D. and Scherphof, G.L. Structure of the cell envelope of Halobacterium halobium. J. Cell. Biol. 71, 1–22 (1970).
Kuschmitz, D. and Hess, B. On the ratio of the proton and photochemical cycles of bacteriorhodopsin. Biochemistry 20, 5950–5957, (1981).
Lozier, R.H., Niederberger, W., Bogomolni, R.N., Hwang, S. and Stoeckenius, W. 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. Biochem. Biophys. Acta, 440, 545–556 (1976).
Govindjee, R., Ebrey, T.G. and Crofts, A.R. The quantum efficiency of proton pumping by the purple membrane of Halobacterium halobium. Biophys. J., 30, 231–242 (1980).
Govndjee, R., Ohno, K. and Ebrey, T.G. Effect of removal of the C-terminal region of bacteriorhodopsin on its light induced H+ changes. Biophys. J. 38, 85–87 (1982).
Gerber, G.E., Wildenauer, D. and Khorana, H.G. Orientation of bacteriorhodopsin in Halobacterium halobium as studied by selective proteolysis. Proc. Natl. Acad. Sci. USA 74, 5426–5430 (1977).
Ovchinnkov, Y.A., Abdulaev, N.G., Feigina, M.Y., Kiselev, A.U. and Lobanov, N.A. Recent fndings in the structure-functional characteristcs of bacteriorhodopsin. FEBS Lett. 84, 1–4 (1977).
Liao, M.-J. and Khorana, H.G. Removal of the carboxyl-terminal peptide does not affect refolding or function of bacteriorhodopsin as a light dependent proton pump J. Biol. Chem. 259, 4194–4199 (1984).
Govindjee, R., Ohno, K., Chang, C.-H. and Ebrey, T.G. In: Transduction in Biological Membranes, Plenum Press, In Press.
Garty, H., Klemperer, G., Eisenbach, M. and Caplan, S.R. FEBS Lett. 81, 238–242 (1977).
Oesterhelt, D. and Stoeckenius, W. Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane, In: Methods in Enzymology (S. Fleischer and L. Packer, eds.), vol. 31, pp. 667–678, New York, Academic Press, (1974).
Oesterhelt, D. Reconstitution of the retinal proteins bacteriorhodopsin and halorhodopsin. In: Methods in Enzymology (L. Packer, ed.), vol. 88, pp. 10–17, New York, Academic Press, (1982).
Mukohata, Y., Sugiyama, Y.,Kaji, Y., Usukura, J. and Yamada, E. The white membrane of crystalline bacterioopsin in Halobacterium halobium strain R1mW and its conversion nto purple membrane by exogenous retinal. Photochem. Photobiol. 33, 593–600, (1981).
Ware, B.R. and Haas, D.D. Electrophoretc light scattering. In: Fast Methods in Physical Biochemistry and Cell Biology (R.I. Sha’afi and S.M. Fernandez,eds.), pp. 174–220, Elsevier Science Publishers, (1983).
Arrio, B., Johannin, G., Carrette, A., Chevallier, J. and Brethes, D. Electrokinetic and hydrodynamic properties of sarcoplasmic reticulum vesicles: a study by laser Doppler electrophoresis and quasielasticlightscattering. Arch. Biochem. Biophys. 228, 220–229, (1984).
Arrio, B., Johannin, G., Volfin, P. and Packer, L. Quas-elastic laser light scatterng and Doppler electrophoresis of purple membranes. Biophys. Soc. Abstr. 45, 212a, (1984).
Arrio, B., Johannin, G., Volfin, P., Lefort-Tran, M., Packer, L., Robinson, A.E. and Hrabeta, E. Aggregation and proton release of purple and white membranes following cleavage of the C-terminal tail of bacteriorhodopsin. Submitted to Archives of Biochemistry and Biophysics (1985).
Wallace, B.A. and Henderson, R. Location of the carboxyl terminus of bacteriorhodopsin in purple membrane. Biophys. J. 39, 233–239, (1982).
Govindgee, R., Ohno, K. and Ebrey, T.G. Effect of the removal of the COOH-terminal region of bacteriorhodopsin on its light-induced H+ changes. Biophys. J. 38, 85–87, (1982).
Walsh, K.A. Trypsinogens and trypsins of various species. Methods in Enzymology (Perlmann,G.E. and Lorand, L.,eds.) vol. 19, pp. 41–63 (1971).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1986 Plenum Press, New York
About this chapter
Cite this chapter
Robinson, A.E., Hrabeta, E., Packer, L. (1986). Measurement of Proton/M412 Ratios in Suspensions of Purple and White Membrane from Halobacterium Halobium. In: Papageorgiou, G.C., Barber, J., Papa, S. (eds) Ion Interactions in Energy Transfer Biomembranes. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-8410-6_17
Download citation
DOI: https://doi.org/10.1007/978-1-4684-8410-6_17
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-8412-0
Online ISBN: 978-1-4684-8410-6
eBook Packages: Springer Book Archive