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. 1976 Jan;125(1):181–190. doi: 10.1128/jb.125.1.181-190.1976

Energy coupling in the active transport of amino acids by bacteriohodopsin-containing cells of Halobacterium holobium.

J S Hubbard, C A Rinehart, R A Baker
PMCID: PMC233350  PMID: 128552

Abstract

Growth of Halobacterium halobium under illumination with limiting aeration induces bacteriorhodopsin formation and renders the cells capable of photophosphorylation. Cells depleted of endogenous reserves by a starvation treatment were used to investigate the means by which energy is coupled to the active transport of [14C]proline, -leucine, and -histidine. Proline was readily accumulated by irradiated cells under anaerobiosis even when the photophosphorylation was abolished by the adenosine triphosphatase inhibitor N,N'-dicyclohexylcarbodimiide (DCCD). The uptake of proline in the dark was limited except when the cells were allowed to accumulate adenosine 5'-triphosphate (ATP) by prior light exposure or by the oxidation of glycerol. DCCD inhibited this dark uptake. These findings essentially support Mitchell's chemiosmotic theory of active transport. The driving force is apparently the proton-motive force developed when protons are extruded from irradiated bacteriorhodopsin or by the dydrolysis of ATP by membrane adenosine triphosphatase. Carbonylcyanide m-chlorophenylhydrazone (CCCP), a proton permeant known to abolish membrane potential, was a strong inhibitor of proline uptake. Leucine transport was also apparently driven by proton-motive force, although its kinetic properties differed from the proline system. Histidine transport is apparently not a chemiosmotic system. Dark- or light-exposed cells show comparable initial rats of histidine uptake, and these processes were only partially inhibited by DCCD or CCCP. The histidine system apparently does not utilize ATP per se since comparable rates of uptake were exhibited by cells of differing intracellular ATP levels. Irradiated cells did effect a greater total accumulation of histidine than dark-exposed cells. These findings suggest that ATP is needed for sustained transport.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altendorf K., Harold F. M., Simoni R. D. Impairment and restoration of the energized state in membrane vesicles of a mutant of Escherichia coli lacking adenosine triphosphatase. J Biol Chem. 1974 Jul 25;249(14):4587–4593. [PubMed] [Google Scholar]
  2. Ames G. F., Lever J. Components of histidine transport: histidine-binding proteins and hisP protein. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1096–1103. doi: 10.1073/pnas.66.4.1096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Asghar S. S., Levin E., Harold F. M. Accumulation of neutral amino acids by Streptococcus faecalis. Energy coupling by a proton-motive force. J Biol Chem. 1973 Aug 10;248(15):5225–5233. [PubMed] [Google Scholar]
  4. Berger E. A. Different mechanisms of energy coupling for the active transport of proline and glutamine in Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1514–1518. doi: 10.1073/pnas.70.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berger E. A., Heppel L. A. Different mechanisms of energy coupling for the shock-sensitive and shock-resistant amino acid permeases of Escherichia coli. J Biol Chem. 1974 Dec 25;249(24):7747–7755. [PubMed] [Google Scholar]
  6. Blaurock A. E., Stoeckenius W. Structure of the purple membrane. Nat New Biol. 1971 Sep 29;233(39):152–155. doi: 10.1038/newbio233152a0. [DOI] [PubMed] [Google Scholar]
  7. Danon A., Stoeckenius W. Photophosphorylation in Halobacterium halobium. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1234–1238. doi: 10.1073/pnas.71.4.1234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harold F. M., Baarda J. R., Baron C., Abrams A. Inhibition of membrane-bound adenosine triphosphatase and of cation transport in Streptococcus faecalis by N,N'-dicyclohexylcarbodiimide. J Biol Chem. 1969 May 10;244(9):2261–2268. [PubMed] [Google Scholar]
  9. Harold F. M. Chemiosmotic interpretation of active transport in bacteria. Ann N Y Acad Sci. 1974 Feb 18;227:297–311. doi: 10.1111/j.1749-6632.1974.tb14395.x. [DOI] [PubMed] [Google Scholar]
  10. Harold F. M. Conservation and transformation of energy by bacterial membranes. Bacteriol Rev. 1972 Jun;36(2):172–230. doi: 10.1128/br.36.2.172-230.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hirata H., Altendorf K., Harold F. M. Energy coupling in membrane vesicles of Escherichia coli. I. Accumulation of metabolites in response to an electrical potential. J Biol Chem. 1974 May 10;249(9):2939–2945. [PubMed] [Google Scholar]
  12. Kaback H. R., Barnes E. M., Jr Mechanisms of active transport in isolated membrane vesicles. II. The mechanism of energy coupling between D-lactic dehydrogenase and beta-galactoside transport in membrane preparations from Escherichia coli. J Biol Chem. 1971 Sep 10;246(17):5523–5531. [PubMed] [Google Scholar]
  13. Kaback H. R., Stadtman E. R. Proline uptake by an isolated cytoplasmic membrane preparation of Escherichia coli. Proc Natl Acad Sci U S A. 1966 Apr;55(4):920–927. doi: 10.1073/pnas.55.4.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kaback H. R. Transport studies in bacterial membrane vesicles. Science. 1974 Dec 6;186(4167):882–892. doi: 10.1126/science.186.4167.882. [DOI] [PubMed] [Google Scholar]
  15. Kanner B. I., Racker E. Light-dependent proton and rubidium translocation in membrane vesicles from Halobacterium halobium. Biochem Biophys Res Commun. 1975 Jan 2;64(3):1054–1061. doi: 10.1016/0006-291x(75)90154-0. [DOI] [PubMed] [Google Scholar]
  16. Kashket E. R., Wilson T. H. Proton-coupled accumulation of galactoside in Streptococcus lactis 7962. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2866–2869. doi: 10.1073/pnas.70.10.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kashket E. R., Wilson T. H. Role of metabolic energy in the transport of -galactosides by Streptococcus lactis. J Bacteriol. 1972 Feb;109(2):784–789. doi: 10.1128/jb.109.2.784-789.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klein W. L., Boyer P. D. Energization of active transport by Escherichia coli. J Biol Chem. 1972 Nov 25;247(22):7257–7265. [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Lombardi F. J., Reeves J. P., Short S. A., Kaback H. R. Evaluation of the chemiosmotic interpretation of active transport in bacterial membrane vesicles. Ann N Y Acad Sci. 1974 Feb 18;227:312–327. doi: 10.1111/j.1749-6632.1974.tb14396.x. [DOI] [PubMed] [Google Scholar]
  21. MacDonald R. E., Lanyi L. K. Light-induced leucine transport in Halobacterium halobium envelope vesicles: a chemiosmotic system. Biochemistry. 1975 Jul;14(13):2882–2889. doi: 10.1021/bi00684a014. [DOI] [PubMed] [Google Scholar]
  22. Mitchell P. Chemiosmotic coupling in energy transduction: a logical development of biochemical knowledge. J Bioenerg. 1972 May;3(1):5–24. doi: 10.1007/BF01515993. [DOI] [PubMed] [Google Scholar]
  23. Mitchell P. Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. Biol Rev Camb Philos Soc. 1966 Aug;41(3):445–502. doi: 10.1111/j.1469-185x.1966.tb01501.x. [DOI] [PubMed] [Google Scholar]
  24. Mitchell P., Moyle J. Acid-base titration across the membrane system of rat-liver mitochondria. Catalysis by uncouplers. Biochem J. 1967 Aug;104(2):588–600. doi: 10.1042/bj1040588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mitchell P. Performance and conservation of osmotic work by proton-coupled solute porter systems. J Bioenerg. 1973 Jan;4(1):63–91. doi: 10.1007/BF01516051. [DOI] [PubMed] [Google Scholar]
  26. ONISHI H., MCCANCE E., GIBBONS N. E. A SYNTHETIC MEDIUM FOR EXTREMELY HALOPHILIC BACTERIA. Can J Microbiol. 1965 Apr;11:365–373. doi: 10.1139/m65-044. [DOI] [PubMed] [Google Scholar]
  27. Oesterhelt D., Hess B. Reversible photolysis of the purple complex in the purple membrane of Halobacterium halobium. Eur J Biochem. 1973 Aug 17;37(2):316–326. doi: 10.1111/j.1432-1033.1973.tb02990.x. [DOI] [PubMed] [Google Scholar]
  28. Oesterhelt D., Stoeckenius W. Functions of a new photoreceptor membrane. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2853–2857. doi: 10.1073/pnas.70.10.2853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pavlasova E., Harold F. M. Energy coupling in the transport of beta-galactosides by Escherichia coli: effect of proton conductors. J Bacteriol. 1969 Apr;98(1):198–204. doi: 10.1128/jb.98.1.198-204.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Racker E., Stoeckenius W. Reconstitution of purple membrane vesicles catalyzing light-driven proton uptake and adenosine triphosphate formation. J Biol Chem. 1974 Jan 25;249(2):662–663. [PubMed] [Google Scholar]
  31. Wilson D. B. Source of energy for the Escherichia coli galactose transport systems induced by galactose. J Bacteriol. 1974 Nov;120(2):866–871. doi: 10.1128/jb.120.2.866-871.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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