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High-field EPR-detected shifts of magnetic tensor components of spin label side chains reveal protein conformational changes: The proton entrance channel of bacteriorhodopsin

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Abstract

Continuous-wave high-field electron paramagnetic resonance (95 GHz, 3.4 T) is performed on a spin label side chain located at residue position 171 in the proton entrance channel of bacterior-hodopsin The conformational differences of three bacteriorhodopsin mutants, the single mutant F171C, the double mutant D96G/F171C, and the triple mutant D96G/F171C/F219L, are reflected in different gxx and Azz tensor component shifts of the nitroxide side chain. The most polar microenvironment is found in the single mutant, whereas the open proton entrance channel reported for the triple mutant allows a reorientation of the nitroxide group towards a microenvironment of lower polarity and/or reduced hydrogen bonding. The experimental data of the double mutant are explained by a light-independent equilibrium of two nitroxide orientations with different polarities of the local microenvironment. Upon illumination the spectrum of the single mutant revealsg xx andA zz tensor component shifts which resemble those determined for the triple mutant in the dark. This result provides strong evidence for a light-induced opening of the proton entrance channel of the single mutant similar to that found in the unilluminated triple mutant, in agreement with electron diffraction data.

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References

  1. Tittor J.: Curr. Biol.1, 534–538 (1991)

    Google Scholar 

  2. Lanyi J.: Biochim. Biophys. Acta1183, 241–261 (1993)

    Article  Google Scholar 

  3. Oesterhelt D.: Curr. Opin. Struct. Biol.8, 489–500 (1998)

    Article  Google Scholar 

  4. Haupts U., Tittor J., Oesterhelt D.: Annu. Rev. Biophys. Biomol. Struct.28, 367–399 (1999)

    Article  Google Scholar 

  5. Váró G., Lanyi J.K.: Biochemistry30, 5008–5015 (1991)

    Article  Google Scholar 

  6. Druckmann S., Friedmann N., Lanyi J.K., Needleman R., Ottolenghi M., Shewes M.: Photochem. Photobiol.56, 1041–1047 (1992)

    Article  Google Scholar 

  7. Hessling B., Herbst J., Rammeisberg R., Gerwert K.: Biophys. J.73, 2071–2080 (1997)

    Article  Google Scholar 

  8. Dencher N.A., Dresselhaus D., Zaccai G., Büldt G.: Proc. Natl. Acad. Sci. USA86, 7876–7879 (1989)

    Article  ADS  Google Scholar 

  9. Koch M.H., Dencher N.A., Oesterhelt D., Plohn H.J., Rapp G., Büldt G.: EMBO J.10, 521–526 (1991)

    Google Scholar 

  10. Subramaniam S., Gerstein M., Oesterhelt D., Henderson R.: EMBO J.12, 1–8 (1993)

    Google Scholar 

  11. Subramaniam S., Lindahl I., Bullough P., Faruqi A.R., Tittor J., Oesterhelt D., Brown L., Lanyi J., Henderson R.: J. Mol. Biol.287, 145–161 (1999)

    Article  Google Scholar 

  12. Subramaniam S., Henderson R.: Nature406, 653–657 (2000)

    Article  ADS  Google Scholar 

  13. Vonck J.: Biochemistry35, 5870–5878 (1996)

    Article  Google Scholar 

  14. Oka T., Kamikubo H., Tokunaga F., Lanyi J.K., Needleman R., Kataoka M.: Biophys. J.76, 1028–1023 (1999)

    Article  Google Scholar 

  15. Sass H.J., Büldt G., Gessenich R., Hehn D., Neff D., Schlesinger R., Berendzen J., Ormos P.: Nature406, 649–652 (2000)

    Article  ADS  Google Scholar 

  16. Hu J.G., Sun B.Q., Bizounok M., Hatcher M.E., Lansing J.C., Raap J., Verdegem P.J., Lugtenburg J., Griffin R.G., Herzfeld J.: Biochemistry37, 8088–8096 (1998)

    Article  Google Scholar 

  17. Moltke S., Wallat I., Sakai N., Nakanishi K., Brown M.F., Heyn M.P.: Biochemistry38, 11762–72 (1999)

    Article  Google Scholar 

  18. Thorgeirsson T.E., Xiao W.Z., Brown L.S., Needleman R., Lanyi J.K., Shin Y.K.: J. Mol. Biol.273, 951–957 (1997)

    Article  Google Scholar 

  19. Rink T., Pfeiffer M., Oesterhelt D., Gerwert K., Steinhoff H.J.: Biophys. J.78, 1519–1530 (2000)

    Article  Google Scholar 

  20. Xiao W., Brown L.S., Needleman R., Lanyi J.K., Shin Y.-K.: J. Mol. Biol.304, 715–721 (2000)

    Article  Google Scholar 

  21. Radzwill N., Gerwert K., Steinhoff H.-J.: Biophys. J.80, 2856–2866 (2001)

    Article  Google Scholar 

  22. Tittor J., Paula S., Subramaniam S., Heberle J., Henderson R., Oesterhelt D.: Biophys. J.78, 478A (2000)

    Google Scholar 

  23. Hubbell W.L., Mchaourab H.S., Altenbach C., Lietzow M.A.: Structure4, 779–783 (1996)

    Article  Google Scholar 

  24. Hubbell W.L., Gross A., Langen R., Lietzow M.A.: Curr. Opin. Struct. Biol.8, 649–656 (1998)

    Article  Google Scholar 

  25. Hubbell W.L., Cafiso D.S., Altenbach C.: Nat. Struct. Biol.7, 735–739 (2000)

    Article  Google Scholar 

  26. Steinhoff H.-J., Savitski A., Wegener C., Pfeiffer M., Plato M., Möbius K.: Biochim. Biophys. Acta1457, 253–262 (2000)

    Article  Google Scholar 

  27. Pfeiffer M., Rink T., Gerwert K., Oesterhelt D., Steinhoff H.J.: J. Mol. Biol.287, 163–171 (1999)

    Article  Google Scholar 

  28. Radionov A.N., Kaulen A.D.: FEBS Lett.387, 122–126 (1996)

    Article  Google Scholar 

  29. Burghaus O., Rohrer M., Gotzinger T., Plato M., Möbius K.: Meas. Sci. Technol.3, 765–774 (1992)

    Article  ADS  Google Scholar 

  30. Prisner T.F., Rohrer M., Möbius K.: Appl. Magn. Reson.7, 167–183 (1994)

    Article  Google Scholar 

  31. Steinhoff H.J., Pfeiffer M., Rink T., Burlon O., Kurz M., Riesle J., Heuberger E., Gerwert K., Oesterhelt D.: Biophys. J.76, 2702–2710 (1999)

    Article  Google Scholar 

  32. Steinhoff H.J., Lieutenant K., Schlitter J.: Z. Naturforsch.44c, 38–46 (1989)

    Google Scholar 

  33. Stone A.J.: Proc. R. Soc. Lond. A271, 424–434 (1963)

    Article  ADS  MATH  Google Scholar 

  34. Burghaus O., Plato M., Rohrer M., Möbius K., MacMillan F., Lubitz W.: J. Phys. Chem.97, 7639–7647 (1993)

    Article  Google Scholar 

  35. Ondar M.A., Grinberg O.Y., Dubinskii A.B., Shestakov A.F., Lebedev Y.S.: Khim. Fiz.2, 54–60 (1983)

    Google Scholar 

  36. Ondar M.A., Grinberg O.Y., Dubinskii A.B., Lebedev Y.S.: Khim. Fiz.3, 527–536 (1984)

    Google Scholar 

  37. Griffith O.H., Dehlinger P.J., Van S.P.: J. Membr. Biol.15, 159–192 (1974)

    Article  Google Scholar 

  38. Kawamura T., Matsunami S., Yonezawa T.: Bull. Chem. Soc. Jpn.40, 1111–1115 (1967)

    Article  Google Scholar 

  39. Earle K.A., Moscicki J.K., Ge M., Budil D.E., Freed J.H.: Biophys. J.66, 1213–1221 (1994)

    Article  Google Scholar 

  40. Gaffney B.J., Marsh D.: Proc. Natl. Acad. Sci. USA95, 12940–12942 (1998)

    Article  ADS  Google Scholar 

  41. Ondar M.A., Grinberg O.Y., Doubinskii A.A., Lebedev Y.S.: Sov. J. Chem. Phys.3, 781–792 (1985)

    Google Scholar 

  42. Engström M., Owenius R., Vahtras O.: Chem. Phys. Lett.338, 407–413 (2001)

    Article  ADS  Google Scholar 

  43. Steinhoff H.J., Müller M., Beier C., Pfeiffer M.: J. Mol. Liq.84, 17–27 (2000)

    Article  Google Scholar 

Download references

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Authors and Affiliations

  1. Max-Planck-Institut für Molekulare Physiologie, Dortmund, Germany

    C. Wegener & H. J. Steinhoff

  2. Institut für Experimentalphysik, Freie Universität Berlin, Berlin, Germany

    A. Savitsky & K. Möbius

  3. Max-Planck-Institut für Biochemie, Martinsried, Germany

    M. Pfeiffer

Authors
  1. C. Wegener
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  2. A. Savitsky
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  3. M. Pfeiffer
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  4. K. Möbius
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  5. H. J. Steinhoff
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Wegener, C., Savitsky, A., Pfeiffer, M. et al. High-field EPR-detected shifts of magnetic tensor components of spin label side chains reveal protein conformational changes: The proton entrance channel of bacteriorhodopsin. Appl. Magn. Reson. 21, 441–452 (2001). https://doi.org/10.1007/BF03162419

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  • DOI: https://doi.org/10.1007/BF03162419

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