Bacterial S-layer protein coupling to lipids: x-ray reflectivity and grazing incidence diffraction studies.
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.
Weygand
M
M
University for Agricultural Sciences, Center for Ultrastructure Research and Ludwig-Boltzmann-Institute for Molecular Nanotechnology, A-1180 Vienna, Austria.
Wetzer
B
B
Pum
D
D
Sleytr
U B
UB
Cuvillier
N
N
Kjaer
K
K
Howes
P B
PB
Lösche
M
M
D000596:Amino Acids / Q000032:analysis; D001407:Bacillus / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D055592:Biophysical Phenomena; D001703:Biophysics; D018360:Crystallography, X-Ray; D008563:Membrane Lipids / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D013057:Spectrum Analysis; D055672:Static Electricity; D014965:X-Rays
D000596:Amino Acids; D001425:Bacterial Outer Membrane Proteins; D008563:Membrane Lipids; D010714:Phosphatidylethanolamines; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine