title: Chemoenzymatic Synthesis of a Phosphorylated Glycoprotein.
abstract: The majority of lysosomal enzymes are targeted to the lysosome by post-translational tagging with N-glycans terminating in mannose-6-phosphate (M6P) residues. Some current enzyme replacement therapies (ERTs) for lysosomal storage disorders are limited in their efficacy by the extent to which the recombinant enzymes bear the M6P-terminated glycans required for effective trafficking. Chemical synthesis was combined with endo-β-N-acetylglucosaminidase (ENGase) catalysis to allow the convergent synthesis of glycosyl amino acids bearing M6P residues. This approach can be extended to the remodeling of proteins, as exemplified by RNase. The powerful synergy of chemical synthesis and ENGase-mediated biocatalysis enabled the first synthesis of a glycoprotein bearing M6P-terminated N-glycans in which the glycans are attached to the peptide backbone by entirely natural linkages.
authors.affiliation: Department of Chemistry, University of Canterbury, Private Bag 4800, Christchurch, 8140, New Zealand.
authors.forename: Pragya
authors.initials: P
authors.lastname: Priyanka
authors.affiliation: Department of Chemistry, Chemistry Research Laboratory, University of Oxford, Mansfield Road, Oxford, OX1 3TA, UK.
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Parsons
authors.affiliation: Callaghan Innovation, School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, 8140, New Zealand.
authors.forename: Antonia
authors.initials: A
authors.lastname: Miller
authors.affiliation: Department of Pharmacology, University of Oxford, Mansfield Road, Oxford, OX1 3QT, UK.
authors.forename: Frances M
authors.initials: FM
authors.lastname: Platt
authors.affiliation: Department of Chemistry, University of Canterbury, Private Bag 4800, Christchurch, 8140, New Zealand. antony.fairbanks@canterbury.ac.nz.
authors.forename: Antony J
authors.identifier: http://orcid.org/0000-0002-9975-3269
authors.initials: AJ
authors.lastname: Fairbanks
mesh_terms: D000118:Acetylglucosaminidase / Q000737:chemistry; D002240:Carbohydrate Sequence; D006023:Glycoproteins / Q000138:chemical synthesis* / Q000737:chemistry; D010766:Phosphorylation
chemical_list: D006023:Glycoproteins; D000118:Acetylglucosaminidase
keywords: ENGase; N-glycans; carbohydrates; glycoproteins; mannose-6-phosphate