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. 1993 Oct;65(4):1449–1459. doi: 10.1016/S0006-3495(93)81214-1

Photochemical conversion of the O-intermediate to 9-cis-retinal-containing products in bacteriorhodopsin films.

A Popp 1, M Wolperdinger 1, N Hampp 1, C Brüchle 1, D Oesterhelt 1
PMCID: PMC1225872  PMID: 8274639

Abstract

The photochemical activity of the O-state was investigated in bacteriorhodopsin (BR) films containing wildtype BR at pH 6.5 in the presence of glycerol. The formation of a photoproduct of O with an absorption maximum at 490 nm and 9-cis-retinal configuration was found. This 490-nm product was named P and shows a slow thermal reaction into a compound with a maximal absorption at 380 nm which was named Q and contains free 9-cis-retinal in the proteins binding site. The photoproducts of O, i.e., P and Q, are very similar, or even identical, to those previously observed in blue membranes. Common to the O-state and blue membrane forms of bacteriorhodopsin is a protonated aspartic acid 85, and we suggest that it is the reduced negative charge around the Schiff base which is responsible for the 9-cis photoisomerization. The release of a proton from aspartic acid 85 is linked to the conversion of the O-state back to the initial state of BR. Therefore the conditions of low proton mobility in BR films containing glycerol favor the accumulation of the O-state. For optical and holographic applications such BR films are very attractive. It is possible to create photoproducts with red light which are thermally stable at room temperature and that can be photochemically erased. Dependent on the light composition both properties can be realized in the same sample material. This feature may bridge the gap between information processing and short-term and long-term storage of information with BR.

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Selected References

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

  1. Bayley H., Huang K. S., Radhakrishnan R., Ross A. H., Takagaki Y., Khorana H. G. Site of attachment of retinal in bacteriorhodopsin. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2225–2229. doi: 10.1073/pnas.78.4.2225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birge R. R. Nature of the primary photochemical events in rhodopsin and bacteriorhodopsin. Biochim Biophys Acta. 1990 Apr 26;1016(3):293–327. doi: 10.1016/0005-2728(90)90163-x. [DOI] [PubMed] [Google Scholar]
  3. Chang C. H., Liu S. Y., Jonas R., Govindjee R. The pink membrane: the stable photoproduct of deionized purple membrane. Biophys J. 1987 Oct;52(4):617–623. doi: 10.1016/S0006-3495(87)83252-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunn R. J., Hackett N. R., McCoy J. M., Chao B. H., Kimura K., Khorana H. G. Structure-function studies on bacteriorhodopsin. I. Expression of the bacterio-opsin gene in Escherichia coli. J Biol Chem. 1987 Jul 5;262(19):9246–9254. [PubMed] [Google Scholar]
  5. Fischer U., Oesterhelt D. Chromophore equilibria in bacteriorhodopsin. Biophys J. 1979 Nov;28(2):211–230. doi: 10.1016/S0006-3495(79)85172-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hampp N., Bräuchle C., Oesterhelt D. Bacteriorhodopsin wildtype and variant aspartate-96 --> aspargine as reversible holographic media. Biophys J. 1990 Jul;58(1):83–93. doi: 10.1016/S0006-3495(90)82355-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harbison G. S., Smith S. O., Pardoen J. A., Winkel C., Lugtenburg J., Herzfeld J., Mathies R., Griffin R. G. Dark-adapted bacteriorhodopsin contains 13-cis, 15-syn and all-trans, 15-anti retinal Schiff bases. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1706–1709. doi: 10.1073/pnas.81.6.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Henderson R., Baldwin J. M., Ceska T. A., Zemlin F., Beckmann E., Downing K. H. Model for the structure of bacteriorhodopsin based on high-resolution electron cryo-microscopy. J Mol Biol. 1990 Jun 20;213(4):899–929. doi: 10.1016/S0022-2836(05)80271-2. [DOI] [PubMed] [Google Scholar]
  9. Hwang S. B., Korenbrot J. I., Stoeckenius W. Transient photovoltages in purple membrane multilayers. Charge displacement in bacteriorhodopsin and its photointermediates. Biochim Biophys Acta. 1978 May 18;509(2):300–317. doi: 10.1016/0005-2736(78)90049-4. [DOI] [PubMed] [Google Scholar]
  10. Kalisky O., Goldschmidt C. R., Ottolenghi M. On the photocycle and light adaptation of dark-adapted bacteriorhodopsin. Biophys J. 1977 Aug;19(2):185–189. doi: 10.1016/S0006-3495(77)85579-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Katre N. V., Wolber P. K., Stoeckenius W., Stroud R. M. Attachment site(s) of retinal in bacteriorhodopsin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4068–4072. doi: 10.1073/pnas.78.7.4068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Korenstein R., Hess B. Hydration effects on cis--trans isomerization of bacteriorhodopsin. FEBS Lett. 1977 Oct 1;82(1):7–11. doi: 10.1016/0014-5793(77)80874-0. [DOI] [PubMed] [Google Scholar]
  13. Kouyama T., Bogomolni R. A., Stoeckenius W. Photoconversion from the light-adapted to the dark-adapted state of bacteriorhodopsin. Biophys J. 1985 Aug;48(2):201–208. doi: 10.1016/S0006-3495(85)83773-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kouyama T., Kinosita K., Jr, Ikegami A. Structure and function of bacteriorhodopsin. Adv Biophys. 1988;24:123–175. doi: 10.1016/0065-227x(88)90006-8. [DOI] [PubMed] [Google Scholar]
  15. Lam E., Fry I., Packer L., Mukohata Y. Comparison of the O640 photo-intermediate and acid-induced species in membrane patches from Halobacterium halobium S9 and R1 mW strains. FEBS Lett. 1982 Sep 6;146(1):106–110. doi: 10.1016/0014-5793(82)80714-x. [DOI] [PubMed] [Google Scholar]
  16. Lanyi J. K., Tittor J., Váró G., Krippahl G., Oesterhelt D. Influence of the size and protonation state of acidic residue 85 on the absorption spectrum and photoreaction of the bacteriorhodopsin chromophore. Biochim Biophys Acta. 1992 Jan 30;1099(1):102–110. [PubMed] [Google Scholar]
  17. Lemke H. D., Oesterhelt D. Lysine 216 is a binding site of the retinyl moiety in bacteriorhodopsin. FEBS Lett. 1981 Jun 15;128(2):255–260. doi: 10.1016/0014-5793(81)80093-2. [DOI] [PubMed] [Google Scholar]
  18. Maeda A., Iwasa T., Yoshizawa T. Formation of 9-cis- and 11-cis-retinal pigments from bacteriorhodopsin by irradiating purple membrane in acid. Biochemistry. 1980 Aug 5;19(16):3825–3831. doi: 10.1021/bi00557a027. [DOI] [PubMed] [Google Scholar]
  19. Metz G., Siebert F., Engelhard M. Asp85 is the only internal aspartic acid that gets protonated in the M intermediate and the purple-to-blue transition of bacteriorhodopsin. A solid-state 13C CP-MAS NMR investigation. FEBS Lett. 1992 Jun 1;303(2-3):237–241. doi: 10.1016/0014-5793(92)80528-o. [DOI] [PubMed] [Google Scholar]
  20. Mowery P. C., Lozier R. H., Chae Q., Tseng Y. W., Taylor M., Stoeckenius W. Effect of acid pH on the absorption spectra and photoreactions of bacteriorhodopsin. Biochemistry. 1979 Sep 18;18(19):4100–4107. doi: 10.1021/bi00586a007. [DOI] [PubMed] [Google Scholar]
  21. Nassal M., Mogi T., Karnik S. S., Khorana H. G. Structure-function studies on bacteriorhodopsin. III. Total synthesis of a gene for bacterio-opsin and its expression in Escherichia coli. J Biol Chem. 1987 Jul 5;262(19):9264–9270. [PubMed] [Google Scholar]
  22. Ni B. F., Chang M., Duschl A., Lanyi J., Needleman R. An efficient system for the synthesis of bacteriorhodopsin in Halobacterium halobium. Gene. 1990 May 31;90(1):169–172. doi: 10.1016/0378-1119(90)90456-2. [DOI] [PubMed] [Google Scholar]
  23. Oesterhelt D., Bräuchle C., Hampp N. Bacteriorhodopsin: a biological material for information processing. Q Rev Biophys. 1991 Nov;24(4):425–478. doi: 10.1017/s0033583500003863. [DOI] [PubMed] [Google Scholar]
  24. Oesterhelt D., Schuhmann L. Reconstitution of bacteriorhodopsin. FEBS Lett. 1974 Aug 30;44(3):262–265. doi: 10.1016/0014-5793(74)81153-1. [DOI] [PubMed] [Google Scholar]
  25. Oesterhelt D., Stoeckenius W. Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol. 1974;31:667–678. doi: 10.1016/0076-6879(74)31072-5. [DOI] [PubMed] [Google Scholar]
  26. Oesterhelt D., Stoeckenius W. Rhodopsin-like protein from the purple membrane of Halobacterium halobium. Nat New Biol. 1971 Sep 29;233(39):149–152. doi: 10.1038/newbio233149a0. [DOI] [PubMed] [Google Scholar]
  27. Otto H., Marti T., Holz M., Mogi T., Stern L. J., Engel F., Khorana H. G., Heyn M. P. Substitution of amino acids Asp-85, Asp-212, and Arg-82 in bacteriorhodopsin affects the proton release phase of the pump and the pK of the Schiff base. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1018–1022. doi: 10.1073/pnas.87.3.1018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Padrós E., Duñach M., Sabés M. Induction of the blue form of bacteriorhodopsin by low concentrations of sodium dodecyl sulfate. Biochim Biophys Acta. 1984 Jan 11;769(1):1–7. doi: 10.1016/0005-2736(84)90002-6. [DOI] [PubMed] [Google Scholar]
  29. Pande C., Callender R. H., Chang C. H., Ebrey T. G. Resonance Raman study of the pink membrane photochemically prepared from the deionized blue membrane of H. halobium. Biophys J. 1986 Sep;50(3):545–549. doi: 10.1016/S0006-3495(86)83493-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Scherrer P., Mathew M. K., Sperling W., Stoeckenius W. Retinal isomer ratio in dark-adapted purple membrane and bacteriorhodopsin monomers. Biochemistry. 1989 Jan 24;28(2):829–834. doi: 10.1021/bi00428a063. [DOI] [PubMed] [Google Scholar]
  31. Soppa J., Oesterhelt D. Bacteriorhodopsin mutants of Halobacterium sp. GRB. I. The 5-bromo-2'-deoxyuridine selection as a method to isolate point mutants in halobacteria. J Biol Chem. 1989 Aug 5;264(22):13043–13048. [PubMed] [Google Scholar]
  32. Soppa J., Otomo J., Straub J., Tittor J., Meessen S., Oesterhelt D. Bacteriorhodopsin mutants of Halobacterium sp. GRB. II. Characterization of mutants. J Biol Chem. 1989 Aug 5;264(22):13049–13056. [PubMed] [Google Scholar]
  33. Subramaniam S., Greenhalgh D. A., Rath P., Rothschild K. J., Khorana H. G. Replacement of leucine-93 by alanine or threonine slows down the decay of the N and O intermediates in the photocycle of bacteriorhodopsin: implications for proton uptake and 13-cis-retinal----all-trans-retinal reisomerization. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6873–6877. doi: 10.1073/pnas.88.15.6873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Subramaniam S., Marti T., Khorana H. G. Protonation state of Asp (Glu)-85 regulates the purple-to-blue transition in bacteriorhodopsin mutants Arg-82----Ala and Asp-85----Glu: the blue form is inactive in proton translocation. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1013–1017. doi: 10.1073/pnas.87.3.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Szundi I., Stoeckenius W. Purple-to-blue transition of bacteriorhodopsin in a neutral lipid environment. Biophys J. 1988 Aug;54(2):227–232. doi: 10.1016/S0006-3495(88)82951-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Szundi I., Stoeckenius W. Surface pH controls purple-to-blue transition of bacteriorhodopsin. A theoretical model of purple membrane surface. Biophys J. 1989 Aug;56(2):369–383. doi: 10.1016/S0006-3495(89)82683-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tolhurst D. J., Lewis P. R. Effect of myelination on the conduction velocity of optic nerve fibres. Ophthalmic Physiol Opt. 1992 Apr;12(2):241–243. doi: 10.1111/j.1475-1313.1992.tb00298.x. [DOI] [PubMed] [Google Scholar]
  38. Váró G., Duschl A., Lanyi J. K. Interconversions of the M, N, and O intermediates in the bacteriorhodopsin photocycle. Biochemistry. 1990 Apr 17;29(15):3798–3804. doi: 10.1021/bi00467a029. [DOI] [PubMed] [Google Scholar]
  39. Váró G., Lanyi J. K. Kinetic and spectroscopic evidence for an irreversible step between deprotonation and reprotonation of the Schiff base in the bacteriorhodopsin photocycle. Biochemistry. 1991 May 21;30(20):5008–5015. doi: 10.1021/bi00234a024. [DOI] [PubMed] [Google Scholar]
  40. Váró G., Lanyi J. K. Photoreactions of bacteriorhodopsin at acid pH. Biophys J. 1989 Dec;56(6):1143–1151. doi: 10.1016/S0006-3495(89)82761-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zimányi L., Lanyi J. K. Iso-halorhodopsin: a stable, 9-cis retinal containing photoproduct of halorhodopsin. Biophys J. 1987 Dec;52(6):1007–1013. doi: 10.1016/S0006-3495(87)83293-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. de Groot H. J., Harbison G. S., Herzfeld J., Griffin R. G. Nuclear magnetic resonance study of the Schiff base in bacteriorhodopsin: counterion effects on the 15N shift anisotropy. Biochemistry. 1989 Apr 18;28(8):3346–3353. doi: 10.1021/bi00434a033. [DOI] [PubMed] [Google Scholar]

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