Bacterial S-layer protein coupling to lipids

X-ray reflectivity and grazing incidence diffraction studies

M. Weygand, B. Wetzer, D. Pum, U.B. Sleytr, N. Cuvillier, K. Kjær, P.B. Howes, M. Lösche

    Research output: Contribution to journalJournal articleResearch

    Abstract

    The coupling of bacterial surface (S)-layer proteins to lipid membranes is studied in molecular detail for proteins from Bacillus sphaericus CCM2177 and B. coagulans E38-66 recrystallized at dipalmitoylphosphatidylethanolamine (DPPE) monolayers on aqueous buffer. A comparison of the monolayer structure before and after protein recrystallization shows minimal reorganization of the lipid chains. By contrast, the lipid headgroups show major rearrangements. For the B. sphaericus CCM2177 protein underneath DPPE monolayers, x-ray reflectivity data suggest that amino acid side chains intercalate the lipid headgroups at least to the phosphate moieties, and probably further beyond. The number of electrons in the headgroup region increases by more than four per lipid. Analysis of the changes of the deduced electron density profiles in terms of a molecular interpretation shows that the phosphatidylethanolamine headgroups must reorient toward the surface normal to accommodate such changes. In terms of the protein structure (which is as yet unknown in three dimensions), the electron density profile reveals a thickness I(z) approximate to 90 Angstrom of the recrystallized S-layer and shows water-filled cavities near its center. The protein volume fraction reaches maxima of >60% in two horizontal sections of the S-layer, close to the lipid monolayer and close to the free subphase. In between it drops to similar to 20%. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of similar to 131 Angstrom.
    Original languageEnglish
    JournalBiophysical Journal
    Volume76
    Issue number1
    Pages (from-to)458-468
    ISSN0006-3495
    Publication statusPublished - 1999

    Cite this

    Weygand, M., Wetzer, B., Pum, D., Sleytr, U. B., Cuvillier, N., Kjær, K., ... Lösche, M. (1999). Bacterial S-layer protein coupling to lipids: X-ray reflectivity and grazing incidence diffraction studies. Biophysical Journal, 76(1), 458-468.
    Weygand, M. ; Wetzer, B. ; Pum, D. ; Sleytr, U.B. ; Cuvillier, N. ; Kjær, K. ; Howes, P.B. ; Lösche, M. / Bacterial S-layer protein coupling to lipids : X-ray reflectivity and grazing incidence diffraction studies. In: Biophysical Journal. 1999 ; Vol. 76, No. 1. pp. 458-468.
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    abstract = "The coupling of bacterial surface (S)-layer proteins to lipid membranes is studied in molecular detail for proteins from Bacillus sphaericus CCM2177 and B. coagulans E38-66 recrystallized at dipalmitoylphosphatidylethanolamine (DPPE) monolayers on aqueous buffer. A comparison of the monolayer structure before and after protein recrystallization shows minimal reorganization of the lipid chains. By contrast, the lipid headgroups show major rearrangements. For the B. sphaericus CCM2177 protein underneath DPPE monolayers, x-ray reflectivity data suggest that amino acid side chains intercalate the lipid headgroups at least to the phosphate moieties, and probably further beyond. The number of electrons in the headgroup region increases by more than four per lipid. Analysis of the changes of the deduced electron density profiles in terms of a molecular interpretation shows that the phosphatidylethanolamine headgroups must reorient toward the surface normal to accommodate such changes. In terms of the protein structure (which is as yet unknown in three dimensions), the electron density profile reveals a thickness I(z) approximate to 90 Angstrom of the recrystallized S-layer and shows water-filled cavities near its center. The protein volume fraction reaches maxima of >60{\%} in two horizontal sections of the S-layer, close to the lipid monolayer and close to the free subphase. In between it drops to similar to 20{\%}. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of similar to 131 Angstrom.",
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    author = "M. Weygand and B. Wetzer and D. Pum and U.B. Sleytr and N. Cuvillier and K. Kj{\ae}r and P.B. Howes and M. L{\"o}sche",
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    Weygand, M, Wetzer, B, Pum, D, Sleytr, UB, Cuvillier, N, Kjær, K, Howes, PB & Lösche, M 1999, 'Bacterial S-layer protein coupling to lipids: X-ray reflectivity and grazing incidence diffraction studies', Biophysical Journal, vol. 76, no. 1, pp. 458-468.

    Bacterial S-layer protein coupling to lipids : X-ray reflectivity and grazing incidence diffraction studies. / Weygand, M.; Wetzer, B.; Pum, D.; Sleytr, U.B.; Cuvillier, N.; Kjær, K.; Howes, P.B.; Lösche, M.

    In: Biophysical Journal, Vol. 76, No. 1, 1999, p. 458-468.

    Research output: Contribution to journalJournal articleResearch

    TY - JOUR

    T1 - Bacterial S-layer protein coupling to lipids

    T2 - X-ray reflectivity and grazing incidence diffraction studies

    AU - Weygand, M.

    AU - Wetzer, B.

    AU - Pum, D.

    AU - Sleytr, U.B.

    AU - Cuvillier, N.

    AU - Kjær, K.

    AU - Howes, P.B.

    AU - Lösche, M.

    PY - 1999

    Y1 - 1999

    N2 - The coupling of bacterial surface (S)-layer proteins to lipid membranes is studied in molecular detail for proteins from Bacillus sphaericus CCM2177 and B. coagulans E38-66 recrystallized at dipalmitoylphosphatidylethanolamine (DPPE) monolayers on aqueous buffer. A comparison of the monolayer structure before and after protein recrystallization shows minimal reorganization of the lipid chains. By contrast, the lipid headgroups show major rearrangements. For the B. sphaericus CCM2177 protein underneath DPPE monolayers, x-ray reflectivity data suggest that amino acid side chains intercalate the lipid headgroups at least to the phosphate moieties, and probably further beyond. The number of electrons in the headgroup region increases by more than four per lipid. Analysis of the changes of the deduced electron density profiles in terms of a molecular interpretation shows that the phosphatidylethanolamine headgroups must reorient toward the surface normal to accommodate such changes. In terms of the protein structure (which is as yet unknown in three dimensions), the electron density profile reveals a thickness I(z) approximate to 90 Angstrom of the recrystallized S-layer and shows water-filled cavities near its center. The protein volume fraction reaches maxima of >60% in two horizontal sections of the S-layer, close to the lipid monolayer and close to the free subphase. In between it drops to similar to 20%. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of similar to 131 Angstrom.

    AB - The coupling of bacterial surface (S)-layer proteins to lipid membranes is studied in molecular detail for proteins from Bacillus sphaericus CCM2177 and B. coagulans E38-66 recrystallized at dipalmitoylphosphatidylethanolamine (DPPE) monolayers on aqueous buffer. A comparison of the monolayer structure before and after protein recrystallization shows minimal reorganization of the lipid chains. By contrast, the lipid headgroups show major rearrangements. For the B. sphaericus CCM2177 protein underneath DPPE monolayers, x-ray reflectivity data suggest that amino acid side chains intercalate the lipid headgroups at least to the phosphate moieties, and probably further beyond. The number of electrons in the headgroup region increases by more than four per lipid. Analysis of the changes of the deduced electron density profiles in terms of a molecular interpretation shows that the phosphatidylethanolamine headgroups must reorient toward the surface normal to accommodate such changes. In terms of the protein structure (which is as yet unknown in three dimensions), the electron density profile reveals a thickness I(z) approximate to 90 Angstrom of the recrystallized S-layer and shows water-filled cavities near its center. The protein volume fraction reaches maxima of >60% in two horizontal sections of the S-layer, close to the lipid monolayer and close to the free subphase. In between it drops to similar to 20%. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of similar to 131 Angstrom.

    KW - Nye funktionelle materialer

    M3 - Journal article

    VL - 76

    SP - 458

    EP - 468

    JO - Biophysical Journal

    JF - Biophysical Journal

    SN - 0006-3495

    IS - 1

    ER -

    Weygand M, Wetzer B, Pum D, Sleytr UB, Cuvillier N, Kjær K et al. Bacterial S-layer protein coupling to lipids: X-ray reflectivity and grazing incidence diffraction studies. Biophysical Journal. 1999;76(1):458-468.