Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1999 Jan;76(1 Pt 1):458–468. doi: 10.1016/S0006-3495(99)77213-9

Bacterial S-layer protein coupling to lipids: x-ray reflectivity and grazing incidence diffraction studies.

M Weygand 1, B Wetzer 1, D Pum 1, U B Sleytr 1, N Cuvillier 1, K Kjaer 1, P B Howes 1, M Lösche 1
PMCID: PMC1302535  PMID: 9876158

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 lz approximately 90 A 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 approximately 20%. Four S-layer protein monomers are located within the unit cell of a square lattice with a spacing of approximately 131 A.

Full Text

The Full Text of this article is available as a PDF (254.7 KB).

Selected References

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

  1. Altmann F. Determination of amino sugars and amino acids in glycoconjugates using precolumn derivatization with o-phthalaldehyde. Anal Biochem. 1992 Jul;204(1):215–219. doi: 10.1016/0003-2697(92)90164-3. [DOI] [PubMed] [Google Scholar]
  2. Amos L. A., Henderson R., Unwin P. N. Three-dimensional structure determination by electron microscopy of two-dimensional crystals. Prog Biophys Mol Biol. 1982;39(3):183–231. doi: 10.1016/0079-6107(83)90017-2. [DOI] [PubMed] [Google Scholar]
  3. Hovmöller S., Sjögren A., Wang D. N. The structure of crystalline bacterial surface layers. Prog Biophys Mol Biol. 1988;51(2):131–163. doi: 10.1016/0079-6107(88)90012-0. [DOI] [PubMed] [Google Scholar]
  4. Küpcü S., Sára M., Sleytr U. B. Liposomes coated with crystalline bacterial cells surface protein (S-layer) as immobilization structures for macromolecules. Biochim Biophys Acta. 1995 May 4;1235(2):263–269. doi: 10.1016/0005-2736(95)80013-6. [DOI] [PubMed] [Google Scholar]
  5. Lösche M., Piepenstock M., Diederich A., Grünewald T., Kjaer K., Vaknin D. Influence of surface chemistry on the structural organization of monomolecular protein layers adsorbed to functionalized aqueous interfaces. Biophys J. 1993 Nov;65(5):2160–2177. doi: 10.1016/S0006-3495(93)81269-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Messner P., Sleytr U. B. Crystalline bacterial cell-surface layers. Adv Microb Physiol. 1992;33:213–275. doi: 10.1016/s0065-2911(08)60218-0. [DOI] [PubMed] [Google Scholar]
  7. Pum D., Sára M., Sleytr U. B. Structure, surface charge, and self-assembly of the S-layer lattice from Bacillus coagulans E38-66. J Bacteriol. 1989 Oct;171(10):5296–5303. doi: 10.1128/jb.171.10.5296-5303.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Pum D., Weinhandl M., Hödl C., Sleytr U. B. Large-scale recrystallization of the S-layer of Bacillus coagulans E38-66 at the air/water interface and on lipid films. J Bacteriol. 1993 May;175(9):2762–2766. doi: 10.1128/jb.175.9.2762-2766.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Schuster B., Pum D., Braha O., Bayley H., Sleytr U. B. Self-assembled alpha-hemolysin pores in an S-layer-supported lipid bilayer. Biochim Biophys Acta. 1998 Mar 13;1370(2):280–288. doi: 10.1016/s0005-2736(97)00274-5. [DOI] [PubMed] [Google Scholar]
  10. Schuster B., Pum D., Sleytr U. B. Voltage clamp studies on S-layer-supported tetraether lipid membranes. Biochim Biophys Acta. 1998 Feb 2;1369(1):51–60. doi: 10.1016/s0005-2736(97)00206-x. [DOI] [PubMed] [Google Scholar]
  11. Sleytr U. B., Sára M. Bacterial and archaeal S-layer proteins: structure-function relationships and their biotechnological applications. Trends Biotechnol. 1997 Jan;15(1):20–26. doi: 10.1016/S0167-7799(96)10063-9. [DOI] [PubMed] [Google Scholar]
  12. Sleytr U. B., Sára M., Küpcü Z., Messner P. Structural and chemical characterization of S-layers of selected strains of Bacillus stearothermophilus and Desulfotomaculum nigrificans. Arch Microbiol. 1986 Oct;146(1):19–24. doi: 10.1007/BF00690152. [DOI] [PubMed] [Google Scholar]
  13. Sára M., Kuen B., Mayer H. F., Mandl F., Schuster K. C., Sleytr U. B. Dynamics in oxygen-induced changes in S-layer protein synthesis from Bacillus stearothermophilus PV72 and the S-layer-deficient variant T5 in continuous culture and studies of the cell wall composition. J Bacteriol. 1996 Apr;178(7):2108–2117. doi: 10.1128/jb.178.7.2108-2117.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sára M., Sleytr U. B. Crystalline bacterial cell surface layers (S-layers): from cell structure to biomimetics. Prog Biophys Mol Biol. 1996;65(1-2):83–111. doi: 10.1016/s0079-6107(96)00007-7. [DOI] [PubMed] [Google Scholar]
  15. Sára M., Sleytr U. B. Relevance of charged groups for the integrity of the S-layer from Bacillus coagulans E38-66 and for molecular interactions. J Bacteriol. 1993 Apr;175(8):2248–2254. doi: 10.1128/jb.175.8.2248-2254.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vaknin D., Kjaer K., Als-Nielsen J., Lösche M. Structural properties of phosphatidylcholine in a monolayer at the air/water interface: Neutron reflection study and reexamination of x-ray reflection measurements. Biophys J. 1991 Jun;59(6):1325–1332. doi: 10.1016/S0006-3495(91)82347-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wetzer B., Pum D., Sleytr U. B. S-layer stabilized solid support lipid bilayers. J Struct Biol. 1997 Jul;119(2):123–128. doi: 10.1006/jsbi.1997.3867. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES