Azide-Functionalized Naphthoxyloside as a Tool for Glycosaminoglycan Investigations.

Daniel Willén,Ulf Ellervik,Roberto Mastio,Sophie Manner,Emil Tykesson,Zackarias Söderlund,Gunilla Westergren-Thorsson

doi:10.1021/acs.bioconjchem.1c00473

Daniel Willén, Ulf Ellervik + Show 5 more

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https://doi.org/10.1021/acs.bioconjchem.1c00473

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Journal: Bioconjugate chemistry	Publication Date: Nov 16, 2021
Citations: 3	License type: CC BY 4.0

Affiliation: Lund University

Abstract

We present a xylosylated naphthoxyloside carrying a terminal azide functionality that can be used for conjugation using click chemistry. We show that this naphthoxyloside serves as a substrate for β4GalT7 and induces the formation of soluble glycosaminoglycan (GAG) chains with physiologically relevant lengths and sulfation patterns. Finally, we demonstrate its usefulness by conjugation to the Alexa Fluor 647 and TAMRA fluorophores and coupling to a surface plasmon resonance chip for interaction studies with the hepatocyte growth factor known to interact with the GAG heparan sulfate.

Highlights

Cellular communication is essential for tissue development, growth, adhesion, coagulation, and pathophysiological processes, for example, tumor development and infection.[1]
As a first attempt toward azide-functionalized naphthoxylosides, we envisioned compounds 5 and 12, where the azide function is connected to the naphthol moiety via a short aliphatic linker
We have shown that XylNapN3 can be taken up by cells and initiate the biosynthesis of soluble GAG chains provided with a handle suitable for functionalization

Summary

Introduction

Cellular communication is essential for tissue development, growth, adhesion, coagulation, and pathophysiological processes, for example, tumor development and infection.[1]. The pattern of alternating disaccharides further classifies the GAGs as heparan sulfate (HS, GlcNAc(β1−4)GlcA(β1−4)) or chondroitin/ dermatan sulfate (CS/DS, GalNAc(β1−4)GlcA(β1−3)). These polymers are postsynthetically sulfated and epimerized to generate a vast structural diversity. The biosynthesis of HS and CS/DS is initiated by xylosylation of a serine residue on the parent protein followed by galactosylation by β-1,4-galactosyltransferase 7 (β4GalT7). This disaccharide is elongated by the addition of another galactose unit and a glucuronic acid moiety to form a common linker region, i.e., GlcA(β1−3)Gal(β1−3)Gal(β1−4)Xyl β (Figure 1A)

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