Protein Aggregation Nucleated by Functionalized Dendritic Polyglycerols.

Dendritic polyglycerols (dPGs) are emerging as important polymers for the study of biological processes due to their relatively low toxicity and excellent biocompatibility. The highly branched nature and high density of endgroups make the dPGs particularly attractive frameworks for the study of multivalent interactions such as multivalent protein-carbohydrate interactions. Here, we report the synthesis of a series of lactose functionalized dPGs with different hydrodynamic radii. A series of lactose functionalized dPGs bearing different densities of lactose functional groups was also synthesized. These lactose functionalized dPGs were used to study the templated aggregation of galectin-3, a galactoside binding protein that is overexpressed during many processes involved in cancer progression. Dynamic light scattering measurements revealed a direct correlation between the hydrodynamic radii of the lactose functionalized dPGs and the size of the galectin-3/lactose functionalized dPG aggregates formed upon mixing the lactose functionalized dPGs with galectin-3 in solution. These studies exposed the critical role of galectin-3's N-terminal domain in formation of galectin-3 multimers and also enabled comparisons of polymer templated aggregation using nonspecific interactions versus specific protein-carbohydrate binding interactions.

Chemoenzymatic Synthesis of Galectin Binding Glycopolymers.

Chemoenzymatic synthesis is an important strategy for the formation of glycopolymers. The use of a smaller number of traditional chemical steps and enzyme catalyzed reactions increases the yield of glycopolymer that can be produced by reducing the overall number of synthetic steps. In addition, chemoenzymatic routes are likely to be more accessible to those without a background in carbohydrate synthesis, making glycopolymers more available for studies across a broader range of scientists. Here, the enzymatic addition of galactose to N-acetylglucosamine functionalized glycodendrimers reduced the requisite number of synthetic steps for the full chemical synthesis of N-acetyl lactosamine (Lac NAc) functionalized dendrimers to four steps. Unpurified cell lysate was used in the enzyme catalyzed glycosylation, and product glycodendrimers were readily purified by dialysis after enzymatic degradation of all protein components of the lysate in the crude reaction mixture. Lac NAc functionalized dendrimers were used very effectively in homotypic cancer cellular aggregation assays and were found to either inhibit or enhance galectin-3 mediated cancer cellular aggregation, with differences in outcomes dependent on the generation of Lac NAc functionalized dendrimers that were used.