Sialic Acid Glycoengineering Using an Unnatural Sialic Acid for the Detection of Sialoglycan Biosynthesis Defects and On-Cell Synthesis of Siglec Ligands

TY - JOUR

T1 - Sialic Acid Glycoengineering Using an Unnatural Sialic Acid for the Detection of Sialoglycan Biosynthesis Defects and On-Cell Synthesis of Siglec Ligands

AU - Büll, Christian

AU - Heise, Torben

AU - Beurskens, Daniëlle M H

AU - Riemersma, Moniek

AU - Ashikov, Angel

AU - Rutjes, Floris P J T

AU - van Kuppevelt, Toin H

AU - Lefeber, Dirk J

AU - den Brok, Martijn H

AU - Adema, Gosse J

AU - Boltje, Thomas J

PY - 2015/10/16

Y1 - 2015/10/16

N2 - 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.

AB - 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.

KW - Animals

KW - Blotting, Western

KW - Carbohydrate Sequence

KW - Female

KW - Flow Cytometry

KW - Glycoproteins/chemistry

KW - Humans

KW - Jurkat Cells

KW - Ligands

KW - Membrane Glycoproteins/chemistry

KW - Mice

KW - Microscopy, Confocal

KW - N-Acetylneuraminic Acid/chemistry

KW - Polysaccharides/chemistry

KW - Protein Binding

KW - Protein Engineering

KW - Sialic Acid Binding Immunoglobulin-like Lectins/chemistry

U2 - 10.1021/acschembio.5b00501

DO - 10.1021/acschembio.5b00501

M3 - Journal article

C2 - 26258433

SN - 1554-8929

VL - 10

SP - 2353

EP - 2363

JO - ACS chemical biology

JF - ACS chemical biology

IS - 10

ER -