Site-specific linking of biomolecules via glycan residues using glycosyltransferases

Abstract

The structural information on glycosyltransferases has revealed that the sugar-donor specificity of these enzymes can be broadened to include modified sugars with a chemical handle that can be utilized for conjugation chemistry. Substitution of Tyr289 to Leu in the catalytic pocket of bovine β-1,4-galactosyltransferase generates a novel glycosyltransferase that can transfer not only Gal but also GalNAc or a C2-modified galactose that has a chemical handle, from the corresponding UDP-derivatives, to the non-reducing end GlcNAc residue of a glycoconjugate. Similarly, the wild-type polypeptide-N-acetyl-galactosaminyltransferase, which naturally transfers GalNAc from UDP-GalNAc, can also transfer C2-modified galactose with a chemical handle from its UDP-derivative to the Ser/Thr residue of a polypeptide acceptor substrate that is tagged as a fusion peptide to a non-glycoprotein. The potential of wild-type and mutant glycosyltransferases to produce glycoconjugates carrying sugar moieties with chemical handle makes it possible to conjugate biomolecules with orthogonal reacting groups at specific sites. This methodology assists in the assembly of bio-nanoparticles that are useful for developing targeted drug-delivery systems and contrast agents for magnetic resonance imaging.