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Relevance of glycosylation of S-layer proteins for cell surface properties.

Schuster B, Sleytr UB - Acta Biomater (2015)

Bottom Line: Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues.The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior.The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices.

View Article: PubMed Central - PubMed

Affiliation: Institute for Synthetic Bioarchitectures, Department of NanoBiotechnology, University of Natural Resources and Life Sciences, Vienna, Muthgasse 11, 1190 Vienna, Austria. Electronic address: bernhard.schuster@boku.ac.at.

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Schematic drawing of the recrystallized S-layer (glyco)proteins SgsE from Geobacillus stearothermophilus NRS 2004/3a. In (A) the recrystallized S-layer glycoprotein wtSgsE (yellow) with branched, fully extended glycan moieties (green) and in (B) the recrystallized S-layer protein rSgsE (yellow) are shown. The intermediary liquid locked-in by the pores within the S-layer lattice and the adjoining, bound water shell is indicated in blue. The scheme is not drawn to scale.
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f0005: Schematic drawing of the recrystallized S-layer (glyco)proteins SgsE from Geobacillus stearothermophilus NRS 2004/3a. In (A) the recrystallized S-layer glycoprotein wtSgsE (yellow) with branched, fully extended glycan moieties (green) and in (B) the recrystallized S-layer protein rSgsE (yellow) are shown. The intermediary liquid locked-in by the pores within the S-layer lattice and the adjoining, bound water shell is indicated in blue. The scheme is not drawn to scale.

Mentions: In the present study, we have exploited for the first time, the relevance of glycosylation at its “native” condition, where a mixture of (glycosylated) S-layer proteins completely covered the surface of the ubiquitous organism G. stearothermophilus NRS 2004/3a, which is not specialized for specific habitats. The recrystallization characteristics and surface properties (i.e., surface hydration, nanoscale fluidic behavior) of this so-called wild-type SgsE (wtSgsE) glycoprotein (schematically depicted in Fig. 1A) are compared to the recombinantly produced protein SgsE (rSgsE), which is N-terminally truncated by 130 amino acids and is lacking the covalently linked carbohydrate moiety (Fig. 1B) [34]. This truncated form has been chosen because (1) the broadest knowledge has accumulated for this non-glycosylated counterpart of wtSgsE, (2) identical S-layer lattice formation compared to wild-type (oblique symmetry, a = 11.6 nm, b = 9.4 nm, and γ ≈ 78° [35]), and (3) highest yield of recombinant protein production [34]. Herein we show that wtSgsE and rSgsE reveal very similar and laterally homogeneous morphology as determined by atomic force microscopy (AFM). Surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with dissipation monitoring (QCM-D) are used to elucidate qualitative differences in the adsorption and self-assembly process, the final mass deposited per unit area, and the coupled and bound water within and on the lattice formed by the glycoprotein wtSgsE and the protein rSgsE, respectively.


Relevance of glycosylation of S-layer proteins for cell surface properties.

Schuster B, Sleytr UB - Acta Biomater (2015)

Schematic drawing of the recrystallized S-layer (glyco)proteins SgsE from Geobacillus stearothermophilus NRS 2004/3a. In (A) the recrystallized S-layer glycoprotein wtSgsE (yellow) with branched, fully extended glycan moieties (green) and in (B) the recrystallized S-layer protein rSgsE (yellow) are shown. The intermediary liquid locked-in by the pores within the S-layer lattice and the adjoining, bound water shell is indicated in blue. The scheme is not drawn to scale.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414373&req=5

f0005: Schematic drawing of the recrystallized S-layer (glyco)proteins SgsE from Geobacillus stearothermophilus NRS 2004/3a. In (A) the recrystallized S-layer glycoprotein wtSgsE (yellow) with branched, fully extended glycan moieties (green) and in (B) the recrystallized S-layer protein rSgsE (yellow) are shown. The intermediary liquid locked-in by the pores within the S-layer lattice and the adjoining, bound water shell is indicated in blue. The scheme is not drawn to scale.
Mentions: In the present study, we have exploited for the first time, the relevance of glycosylation at its “native” condition, where a mixture of (glycosylated) S-layer proteins completely covered the surface of the ubiquitous organism G. stearothermophilus NRS 2004/3a, which is not specialized for specific habitats. The recrystallization characteristics and surface properties (i.e., surface hydration, nanoscale fluidic behavior) of this so-called wild-type SgsE (wtSgsE) glycoprotein (schematically depicted in Fig. 1A) are compared to the recombinantly produced protein SgsE (rSgsE), which is N-terminally truncated by 130 amino acids and is lacking the covalently linked carbohydrate moiety (Fig. 1B) [34]. This truncated form has been chosen because (1) the broadest knowledge has accumulated for this non-glycosylated counterpart of wtSgsE, (2) identical S-layer lattice formation compared to wild-type (oblique symmetry, a = 11.6 nm, b = 9.4 nm, and γ ≈ 78° [35]), and (3) highest yield of recombinant protein production [34]. Herein we show that wtSgsE and rSgsE reveal very similar and laterally homogeneous morphology as determined by atomic force microscopy (AFM). Surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with dissipation monitoring (QCM-D) are used to elucidate qualitative differences in the adsorption and self-assembly process, the final mass deposited per unit area, and the coupled and bound water within and on the lattice formed by the glycoprotein wtSgsE and the protein rSgsE, respectively.

Bottom Line: Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues.The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior.The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices.

View Article: PubMed Central - PubMed

Affiliation: Institute for Synthetic Bioarchitectures, Department of NanoBiotechnology, University of Natural Resources and Life Sciences, Vienna, Muthgasse 11, 1190 Vienna, Austria. Electronic address: bernhard.schuster@boku.ac.at.

Show MeSH
Related in: MedlinePlus