title: Sialic Acid Glycoengineering Using an Unnatural Sialic Acid for the Detection of Sialoglycan Biosynthesis Defects and On-Cell Synthesis of Siglec Ligands.
abstract: Sialoglycans play a vital role in physiology, and aberrant sialoglycan expression is associated with a broad spectrum of diseases. Since biosynthesis of sialoglycans is only partially regulated at the genetic level, chemical tools are crucial to study their function. Here, we report the development of propargyloxycarbonyl sialic acid (Ac5NeuNPoc) as a powerful tool for sialic acid glycoengineering. Ac5NeuNPoc showed strongly increased labeling efficiency and exhibited less toxicity compared to those of widely used mannosamine analogues in vitro and was also more efficiently incorporated into sialoglycans in vivo. Unlike mannosamine analogues, Ac5NeuNPoc was exclusively utilized in the sialoglycan biosynthesis pathway, allowing a genetic defect in sialic acid biosynthesis to be specifically detected. Furthermore, Ac5NeuNPoc-based sialic acid glycoengineering enabled the on-cell synthesis of high-affinity Siglec-7 ligands and the identification of a novel Siglec-2 ligand. Thus, Ac5NeuNPoc glycoengineering is a highly efficient, nontoxic, and selective approach to study and modulate sialoglycan interactions on living cells.
authors.forename: Christian
authors.initials: C
authors.lastname: Büll
authors.affiliation: Cluster for Molecular Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen , Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors.forename: Torben
authors.initials: T
authors.lastname: Heise
authors.forename: Daniëlle M H
authors.initials: DM
authors.lastname: Beurskens
authors.forename: Moniek
authors.initials: M
authors.lastname: Riemersma
authors.forename: Angel
authors.initials: A
authors.lastname: Ashikov
authors.affiliation: Cluster for Molecular Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen , Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors.forename: Floris P J T
authors.initials: FP
authors.lastname: Rutjes
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.forename: Dirk J
authors.initials: DJ
authors.lastname: Lefeber
authors.forename: Martijn H
authors.initials: MH
authors.lastname: den Brok
authors.forename: Gosse J
authors.initials: GJ
authors.lastname: Adema
authors.affiliation: Cluster for Molecular Chemistry, Institute for Molecules and Materials, Radboud University Nijmegen , Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Boltje
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D005260:Female; D005434:Flow Cytometry; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D008562:Membrane Glycoproteins / Q000737:chemistry; D051379:Mice; D018613:Microscopy, Confocal; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D011485:Protein Binding; D015202:Protein Engineering; D063265:Sialic Acid Binding Immunoglobulin-like Lectins / Q000737:chemistry*
chemical_list: D006023:Glycoproteins; D008024:Ligands; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; D019158:N-Acetylneuraminic Acid