Streamlining the chemoenzymatic synthesis of complex N-glycans by a stop and go strategy

Nat Chem. 2019 Feb;11(2):161-169. doi: 10.1038/s41557-018-0188-3. Epub 2018 Dec 10.

Abstract

Contemporary chemoenzymatic approaches can provide highly complex multi-antennary N-linked glycans. These procedures are, however, very demanding and typically involve as many as 100 chemical steps to prepare advanced intermediates that can be diversified by glycosyltransferases in a branch-selective manner to give asymmetrical structures commonly found in nature. Only highly specialized laboratories can perform such syntheses, which greatly hampers progress in glycoscience. Here we describe a biomimetic approach in which a readily available bi-antennary glycopeptide can be converted in ten or fewer chemical and enzymatic steps into multi-antennary N-glycans that at each arm can be uniquely extended by glycosyltransferases to give access to highly complex asymmetrically branched N-glycans. A key feature of our approach is the installation of additional branching points using recombinant MGAT4 and MGAT5 in combination with unnatural sugar donors. At an appropriate point in the enzymatic synthesis, the unnatural monosaccharides can be converted into their natural counterpart, allowing each arm to be elaborated into a unique appendage.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Asparagine / metabolism
  • Biomimetic Materials / metabolism*
  • Carbohydrate Sequence
  • Escherichia coli / enzymology
  • Escherichia coli Proteins / metabolism
  • Galactosidases / metabolism
  • Glycopeptides / metabolism
  • Glycosylation
  • N-Acetylglucosaminyltransferases / metabolism
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism*
  • Sialyltransferases / metabolism

Substances

  • Escherichia coli Proteins
  • Glycopeptides
  • Polysaccharides
  • Asparagine
  • N-Acetylglucosaminyltransferases
  • Sialyltransferases
  • Galactosidases