Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1981 Aug;317:347–364. doi: 10.1113/jphysiol.1981.sp013829

Effect of pH on the formation and decay of the metarhodopsins of the frog.

C Baumann, W Zeppenfeld
PMCID: PMC1246793  PMID: 6975819

Abstract

1. Suspensions of membrane vesicles were prepared by sonication of dark-adapted frog rod outer segments. The pH was adjusted to one of twelve values between 5.0 and 9.8; the temperature was 15 degrees C. Each preparation was exposed to a single yellow flash of 120 microseconds duration, and rapid slow changes of absorbance were measured at 475 nm wavelength. 2. Rapid changes consist of a transient rise followed by a diphasic decay to a new level of absorbance which is lower than that before the light exposure. The level of absorbance reached at the end of the rapid changes is lower at lower pH. 3. Kinetic analysis reveals that three reactants take part in the rapid reactions, viz. metarhodopsin I and two isochromic forms of metarhodopsin II named metarhodopsin II' and metarhodopsin II". The kinetics of the reversible transition from metarhodopsin I to metarhodopsin II' are not measurably influenced by the pH of the medium. However, the reversible reaction of metarhodopsin II' with metarhodopsin II" is dependent on pH because metarhodopsin II" is encountered either as a protonated or as a deprotonated compound. 4. Slow reactions are due to metarhodopsin I and to metarhodopsin III. A combined quantitative analysis of both rapid and slow reactions involves a reversible reaction between metarhodopsin II' and metarhodopsin III and a hydrolysis of metarhodopsin II' to retinal and opsin. The scheme employed accounts for the pH dependence of all equilibria although only one of the metarhodopsins reacts directly with protons.

Full text

PDF
347

Selected References

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

  1. Adams A. J., Tanaka M., Shichi H. Concanavalin A binding to rod outer segment membranes: usefulness for preparation of intact disks. Exp Eye Res. 1978 Nov;27(5):595–605. doi: 10.1016/0014-4835(78)90144-6. [DOI] [PubMed] [Google Scholar]
  2. Baumann C. Kinetics of slow thermal reactions during the bleaching of rhodopsin in the perfused frog retina. J Physiol. 1972 May;222(3):643–663. doi: 10.1113/jphysiol.1972.sp009819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baumann C. The equilibrium between metarhodopsin I and metarhodopsin II in the isolated frog retina. J Physiol. 1978 Jun;279:71–80. doi: 10.1113/jphysiol.1978.sp012331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumann C. The formation of metarhodospin380 in the retinal rods of the frog. J Physiol. 1976 Jul;259(2):357–366. doi: 10.1113/jphysiol.1976.sp011470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bowmaker J. K. The photoproducts of retinal-based visual pigments in situ: a contrast between Rana pipiens and Gekko gekko. Vision Res. 1973 Jul;13(7):1227–1240. doi: 10.1016/0042-6989(73)90199-5. [DOI] [PubMed] [Google Scholar]
  6. Chabre M., Breton J. The orientation of the chromophore of vertebrate rhodopsin in the "meta" intermediate states and the reversibility of the meta II-meta III transition. Vision Res. 1979;19(9):1005–1018. doi: 10.1016/0042-6989(79)90226-8. [DOI] [PubMed] [Google Scholar]
  7. Donner K. O., Hemilä S. Kinetics of long-lived rhodopsin photoproducts in the frog retina as a function of the amount bleached. Vision Res. 1975 Aug-Sep;15:985–995. doi: 10.1016/0042-6989(75)90241-2. [DOI] [PubMed] [Google Scholar]
  8. Emrich H. M., Reich R. Uber Primärreaktionen beim Schvorgang. Thermodynamischer und kinetischer Einfluss des pH-Wertes auf die Metarhodopsin-I-II-Umwandlung. Protonenverbrauch als Auswirkung einer Konformationsänderung. Z Naturforsch C. 1974 Sep-Oct;29C(9-10):577–591. [PubMed] [Google Scholar]
  9. Ernst W., Kemp C. M., White H. A. Studies on the effects of bleaching amphibian rod pigments in situ II. the kinetics of the slow bleaching reactions in axolotl red rods. Exp Eye Res. 1978 Mar;26(3):337–350. doi: 10.1016/0014-4835(78)90080-5. [DOI] [PubMed] [Google Scholar]
  10. Falk G., Fatt P. Rapid hydrogen ion uptake of rod outer segments and rhodopsin solutions on illumination. J Physiol. 1966 Mar;183(1):211–224. doi: 10.1113/jphysiol.1966.sp007861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HUBBARD R., BROWN P. K., KROPF A. Vertebrate lumi- and meta-rhodopsins. Nature. 1959 Feb 14;183(4659):442–446. [PubMed] [Google Scholar]
  12. Hagins W. A., Yoshikami S. Ionic mechanisms in excitation of photoreceptors. Ann N Y Acad Sci. 1975 Dec 30;264:314–325. doi: 10.1111/j.1749-6632.1975.tb31492.x. [DOI] [PubMed] [Google Scholar]
  13. Hubbell W. L., Bownds M. D. Visual transduction in vertebrate photoreceptors. Annu Rev Neurosci. 1979;2:17–34. doi: 10.1146/annurev.ne.02.030179.000313. [DOI] [PubMed] [Google Scholar]
  14. McConnell D. G., Rafferty C. N., Dilley R. A. The light-induced proton uptake in bovine retinal outer segment fragments. J Biol Chem. 1968 Nov 25;243(22):5820–5826. [PubMed] [Google Scholar]
  15. Nakano T., Ikai A., Nishigai M., Noda H. Orientation of the rhodopsin sugar moiety in bovine disk membrane. J Biochem. 1979 May;85(5):1339–1346. [PubMed] [Google Scholar]
  16. Ostroy S. E. Hydrogen ion changes of rhodopsin. pK changes and the thermal decay of metarhodopsin II380. Arch Biochem Biophys. 1974 Sep;164(1):275–284. doi: 10.1016/0003-9861(74)90032-0. [DOI] [PubMed] [Google Scholar]
  17. RADDING C. M., WALD G. The stability of rhodopsin and opsin; effects of pH and aging. J Gen Physiol. 1956 Jul 20;39(6):923–933. doi: 10.1085/jgp.39.6.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rahn H. Evolution of the gas transport system in vertebrates. Proc R Soc Med. 1966 Jun;59(6):493–494. doi: 10.1177/003591576605900608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Reuter T. Photoregeneration of rhodopsin and isorhodopsin from metarhodopsin III in the frog retina. Vision Res. 1976;16(9):909–917. doi: 10.1016/0042-6989(76)90220-0. [DOI] [PubMed] [Google Scholar]
  20. WULFF V. J., ADAMS R. G., LINSCHITZ H., ABRAHAMSON E. W. Effect of flash illumination on rhodopsin in solution. Ann N Y Acad Sci. 1959 Nov 12;74(2):281–290. doi: 10.1111/j.1749-6632.1958.tb39551.x. [DOI] [PubMed] [Google Scholar]
  21. Walker J. L., Brown H. M. Intracellular ionic activity measurements in nerve and muscle. Physiol Rev. 1977 Oct;57(4):729–778. doi: 10.1152/physrev.1977.57.4.729. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES