Monodisperse protein-based glycopolymers via a combined biosynthetic and chemical approach.

Helical protein polymers with sequences comprising primarily alanine and glutamine have been designed to contain glutamic acid residues at distances that are targeted to match the receptor spacing of certain toxins and lectins. These proteins are readily expressed and purified from E. coli and are highly helical under a variety of solution conditions. The helical artificial proteins are also competent for chemical modification with saccharides for inhibition of select bacterial toxins and lectins. In the investigations reported here, multivalent artificial glycoproteins bearing galactose moieties as pendant groups have been prepared via the coupling reaction of amine-functionalized galactose with the glutamic acid functional groups of the protein polymer. Glycosylation of proteins was confirmed via mass spectrometry, NMR spectroscopy, SDS-PAGE, and photometric methods. CD spectroscopy shows that the resulting glycosylated proteins maintain a highly helical structure, and competitive ELISA assays suggest the efficient binding of these glycoproteins to cholera toxin. These results demonstrate that the integration of biological and chemical approaches in the synthesis of well-defined polymeric structures offers significant opportunities in the purposeful design of glycopolymers for applications in biology.