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Combination of extended X-ray absorption fine structure spectroscopy with lipidic cubic phases for the study of cation binding in bacteriorhodopsin

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Abstract

We have performed a quantitative X-ray absorption fine structure analysis of bacteriorhodopsin in purple membrane patches and in lipidic cubic phases regenerated with Mn2+. Lipidic cubic phases and purple membrane results have been compared, demonstrating that the lipidic cubic phase process does not introduce relevant distortions in the local geometry of the cation binding sites. For both samples, we have observed similarities for Mn2+ coordination in terms of type, number, and average distances of surrounding atoms, indicating a first coordination shell composed by 6 O atoms, and 3/4 C atoms located in the second coordination shell.

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References

  • Ankudinov AL, Ravel B, Rehr JJ, Conradson SD (1998) Real-space multiple-scattering calculation and interpretation of X-ray-absorption near edge structure. Phys Rev B 58:7565–7576

    Article  CAS  Google Scholar 

  • Belrhali H, Nollert P, Royant A, Menzel C, Rosenbusch JP, Landau EM, Pebay-Peyroula E (1999) Protein, lipid and water organization in bacteriorhodopsin crystals: a molecular view of the purple membrane at 1.9 A resolution. Structure 7(8):909–917

    Article  PubMed  CAS  Google Scholar 

  • Dracheva S, Bose S, Hendler RW (1996) Chemical and functional studies on the importance of purple membrane lipids in bacteriorhodopsin. Photocycle Behav 382(1–2):209–212

    CAS  Google Scholar 

  • Duñach M, Seigneuret M, Rigaud JL, Padrós E (1986) The relationship between the chromophore moiety and the cation binding sites in bacteriorhodopsin. Biosci Rep 6(11):961–966

    Article  PubMed  Google Scholar 

  • Duñach M, Seigneuret M, Riguad JL, Padrós E (1987) Characterization of the cation binding-sites of the purple membrane—electron-spin-resonance and flash-photolysis studies. Biochemistry 26(4):1179–1186

    Article  Google Scholar 

  • Duñach M, Padrós E, Seigneuret M, Rigaud JL (1988a) On the molecular mechanism of the blue to purple transition of bacteriorhodopsin. UV-difference spectroscopy and electron spin resonance studies. J Biol Chem 263(16):7555–7559

    PubMed  Google Scholar 

  • Duñach M, Seigneuret M, Rigaud JL, Padrós E (1988b) Influence of cations on the blue to purple transition of bacteriorhodopsin. Comparison of Ca2+ and Hg2+ binding and their effect on the surface potential. J Biol Chem 263(33):17378–17384

    PubMed  Google Scholar 

  • Duñach M, Padrós E, Muga A, Arrondo JL (1989) Fourier-transform infrared studies on cation binding to native and modified purple membranes. Biochemistry 28(22):8940–8945

    Article  PubMed  Google Scholar 

  • Eliash T, Weiner L, Ottolenghi M, Sheves M (2001) Specific binding sites for cations in bacteriorhodopsin. Biophys J 81(2):1155–1162

    Article  PubMed  CAS  Google Scholar 

  • Facciotti MT, Rouhani S, Burkard FT, Betancourt FM, Downing KH, Rose RB, McDermott G, Glaeser RM (2001) Structure of an early intermediate in the M-state phase of the bacteriorhodopsin photocycle. Biophys J 81(6):3442–3455

    Article  PubMed  CAS  Google Scholar 

  • Hirai T, Subramaniam S (2003) Structural insights into the mechanism of proton pumping by bacteriorhodopsin. FEBS Lett 545(1):2–8

    Article  PubMed  CAS  Google Scholar 

  • Jonas R, Ebrey TG (1991) Binding of a single divalent cation directly correlates with the blue-to-purple transition in bacteriorhodopsin. Proc Nat Acad Sci USA 88(1):149–153

    Article  PubMed  CAS  Google Scholar 

  • Kimura Y, Ikegami A, Stoeckenius W (1984) Salt and pH-dependent changes of the purple membrane absorption spectrum. Photochem Photobiol 40(5):641–646

    Article  PubMed  CAS  Google Scholar 

  • Lanyi JK (2001) X-ray crystallography of bacteriorhodopsin and its photointermediates: insights into the mechanism of proton transport. Biochemistry (Moscow) 66(11):1192–1196

    Article  CAS  Google Scholar 

  • Luecke H, Schobert B, Richter HT, Cartailler JP, Lanyi JK (1999) Structure of bacteriorhodopsin at 1.55 A resolution. J Mol Biol 291(4):899–911. doi:10.1006/jmbi.1999.3027

    Article  PubMed  CAS  Google Scholar 

  • Newville M (2001) IFEFFIT: interactive XAFS analysis and FEFF fitting. J Synchrotron Radiat 8:3

    Google Scholar 

  • Nollert P, Royant A, Pebay-Peyroula E, Landau EM (1999) Detergent-free membrane protein crystallization. FEBS Lett 457(2):205–208

    Article  PubMed  CAS  Google Scholar 

  • Oesterhelt D, Stoeckenius W (1974) Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol 31(Pt A):667–678

    Article  PubMed  CAS  Google Scholar 

  • Ravel B, Newville M (2005) ATHENA, ARTHEMIS, HEPHAESTUS: data analysis for x-ray absorption spectroscopy using IFEFFIT. J Sybchrotron Radiat 12:537–541

    Article  CAS  Google Scholar 

  • Sanz C, Marquez M, Perálvarez A, Elouatik S, Sepulcre F, Querol E, Lazarova T, Padrós E (2001) Contribution of extracellular Glu residues to the structure and function of bacteriorhodopsin. Presence of specific cation-binding sites. J Biol Chem 276(44):40788–40794

    Article  PubMed  CAS  Google Scholar 

  • Sepulcre F, Cladera J, Garcia J, Proietti MG, Torres J, Padrós E (1996) An extended x-ray absorption fine structure study of the high-affinity cation-binding site in the purple membrane. Biophys J 70(2):852–856

    Article  PubMed  CAS  Google Scholar 

  • Sepulcre F, Proietti MG, Benfatto M, Della Longa S, Garcia J, Padrós E (2004) A quantitative XANES analysis of the calcium high-affinity binding site of the purple membrane. Biophys J 87(1):513–520

    Article  PubMed  CAS  Google Scholar 

  • Sepulcre F, Proietti M, Benfatto M, Della Longa S, Garcia J, Padrós E (2005) An XAS study of the cation binding sites in the purple membrane of Halobacterium salinarum. Physica Scripta T115:855–858

    Article  CAS  Google Scholar 

  • Sepulcre F, Cordomi A, Proietti MG, Perez JJ, Garcia J, Querol E, Padrós E (2007) X-ray absorption and molecular dynamics study of cation binding sites in the purple membrane. Proteins 67(2):360–374

    Article  PubMed  CAS  Google Scholar 

  • Sevillano E, Meuth H, Rehr JJ (1979) Extended X-Ray Absorption Fine-Structure Debye-Waller Factors. 1 Monatomic crystals. Phys Rev B 20(12):4908–4911

    Article  CAS  Google Scholar 

  • Tuzi S, Yamaguchi S, Tanio M, Konishi H, Inoue S, Naito A, Needleman R, Lanyi JK, SaitÃ’ H (1999) Location of a cation-binding site in the loop between helices F and G of bacteriorhodopsin as studied by 13C NMR. Biophys J 76(3):1523–1531

    Article  PubMed  CAS  Google Scholar 

  • Wassenaar TA, Daura X, Padrós E, Mark AE (2009) Calcium binding to the purple membrane: A molecular dynamics study. Prot-Struct Func Bioinfo 74(3):669–681

    Article  CAS  Google Scholar 

  • Yang D, el-Sayed MA (1995) The Ca2+ binding to deionized monomerized and to retinal removed bacteriorhodopsin. Biophys J 69(5):2056–2059

    Article  PubMed  CAS  Google Scholar 

  • Yoo SK, Awad ES, Elsayed MA (1995) Comparison between the Binding of Ca2+ and Mg2+ to the 2 High-Affinity Sites of Bacteriorhodopsin. J Phys Chem 99(29):11600–11604

    Article  CAS  Google Scholar 

  • Zhang NY, el-Sayed MA (1993) The C-terminus and the Ca2+ low-affinity binding sites in bacteriorhodopsin. Biochemistry 32(51):14173–14175

    Article  PubMed  CAS  Google Scholar 

  • Zhang YN, Sweetman LL, Awad ES, El-Sayed MA (1992) Nature of the individual Ca binding sites in Ca-regenerated bacteriorhodopsin. Biophys J 61(5):1201–1206

    Article  PubMed  CAS  Google Scholar 

  • Zhang YN, el-Sayed MA, Bonet ML, Lanyi JK, Chang M, Ni B, Needleman R (1993) Effects of genetic replacements of charged and H-bonding residues in the retinal pocket on Ca2+ binding to deionized bacteriorhodopsin. Proc Natl Acad Sci USA 90(4):1445–1449

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We acknowledge the European Synchrotron Radiation Facility for provision of synchrotron radiation facilities and we would like to thank Dr. Pieter Glatzel for assistance in using the ID26 beam line. This work was supported by a Dirección General de Investigación grant BFU2006-04656/BMC, BFU2009-08758, and a Marie Curie International Outgoing Fellowship to A.P.-M. within the 7th European Community Framework Programme (PIOF-GA-2009-237120).

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Correspondence to Alex Perálvarez-Marín or Francesc Sepulcre.

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A. Perálvarez-Marín and F. Sepulcre contributed equally.

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Perálvarez-Marín, A., Sepulcre, F., Márquez, M. et al. Combination of extended X-ray absorption fine structure spectroscopy with lipidic cubic phases for the study of cation binding in bacteriorhodopsin. Eur Biophys J 40, 1007–1012 (2011). https://doi.org/10.1007/s00249-011-0714-3

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  • DOI: https://doi.org/10.1007/s00249-011-0714-3

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