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Recombinant Glycans on an S‐Layer Self‐Assembly Protein: A New Dimension for Nanopatterned Biomaterials
Author(s) -
Steiner Kerstin,
Hanreich Angelika,
Kainz Birgit,
Hitchen Paul G.,
Dell Anne,
Messner Paul,
Schäffer Christina
Publication year - 2008
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.200701215
Subject(s) - nanotechnology , self assembly , glycan , layer (electronics) , recombinant dna , dimension (graph theory) , materials science , chemistry , biochemistry , glycoprotein , gene , mathematics , pure mathematics
Crucial biological phenomena are mediated through carbohydrates that are displayed in a defined manner and interact with molecular scale precision. We lay the groundwork for the integration of recombinant carbohydrates into a “biomolecular construction kit” for the design of new biomaterials, by utilizing the self‐assembly system of the crystalline cell surface (S)‐layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a. SgsE is a naturally O ‐glycosylated protein, with intrinsic properties that allow it to function as a nanopatterned matrix for the periodic display of glycans. By using a combined carbohydrate/protein engineering approach, two types of S‐layer neoglycoproteins are produced in Escherichia coli . Based on the identification of a suitable periplasmic targeting system for the SgsE self‐assembly protein as a cellular prerequisite for protein glycosylation, and on engineering of one of the natural protein O ‐glycosylation sites into a target for N‐glycosylation, the heptasaccharide from the AcrA protein of Campylobacter jejuni and the O7 polysaccharide of E. coli are co‐ or post‐translationally transferred to the S‐layer protein by the action of the oligosaccharyltransferase PglB. The degree of glycosylation of the S‐layer neoglycoproteins after purification from the periplasmic fraction reaches completeness. Electron microscopy reveals that recombinant glycosylation is fully compatible with the S‐layer protein self‐assembly system. Tailor‐made (“functional”) nanopatterned, self‐assembling neo glycoproteins may open up new strategies for influencing and controlling complex biological systems with potential applications in the areas of biomimetics, drug targeting, vaccine design, or diagnostics.