Engineering the sialome of mammalian cells with sialic acid mimetics

Daniël L A H Hornikx; Eline A Visser; Venetia Psomiadou; Christian Büll; Thomas J Boltje

doi:10.1016/j.xpro.2023.102330

Engineering the sialome of mammalian cells with sialic acid mimetics

STAR Protoc. 2023 Sep 15;4(3):102330. doi: 10.1016/j.xpro.2023.102330. Epub 2023 Jun 28.

Authors

Daniël L A H Hornikx¹, Eline A Visser², Venetia Psomiadou², Christian Büll³, Thomas J Boltje⁴

Affiliations

¹ Department of Biomolecular Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen, the Netherlands.
² Department of Synthetic Organic Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen, the Netherlands.
³ Department of Biomolecular Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen, the Netherlands. Electronic address: [email protected].
⁴ Department of Synthetic Organic Chemistry, Institute for Molecules and Materials, Radboud University, Nijmegen, the Netherlands. Electronic address: [email protected].

Abstract

Mammalian glycans show a diversity in sialic acid capping, constituting the sialome. Sialic acids can be extensively modified chemically, yielding sialic acid mimetics (SAMs). Here, we present a protocol for detecting and quantifying incorporative SAMs using microscopy and flow cytometry, respectively. We detail steps for linking SAMS to proteins with western blotting. Lastly, we detail procedures for incorporative or inhibitory SAMs and how SAMs can be used for the on-cell synthesis of high-affinity Siglec ligands. For complete details on the use and execution of this protocol, please refer to Büll et al.¹ and Moons et al.².

Keywords: Cell culture; Chemistry; Flow Cytometry/Mass Cytometry.

MeSH terms

Animals
Flow Cytometry
Ligands
Mammals / metabolism
N-Acetylneuraminic Acid* / metabolism
Sialic Acid Binding Immunoglobulin-like Lectins* / metabolism
Sialic Acids / metabolism

Substances

Ligands
N-Acetylneuraminic Acid
Sialic Acid Binding Immunoglobulin-like Lectins
Sialic Acids