Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Oct 19;10(63):38561-38565.
doi: 10.1039/d0ra08400j. eCollection 2020 Oct 15.

Synthesis of aryl azide chain-end functionalized N-linked glycan polymers and their photo-labelling of specific protein

Ka Keung Chan  1 , Qiaoshi Lei  2 , Jinshan Tang  2 , Xue-Long Sun  1
Affiliations

Synthesis of aryl azide chain-end functionalized N-linked glycan polymers and their photo-labelling of specific protein

Ka Keung Chan et al. RSC Adv. .

Abstract

We report a straightforward synthesis of aryl azide chain-end functionalized N-linked glycan polymers and its application for affinity-assisted photo-labelling of specific protein. The aryl azide chain-end functionalized N-glycan polymers, including N-galactosyl, N-glucosyl, and N-lactosyl polymer, were synthesized from free glycan via glycosylamine intermediates followed by acrylation and polymerization via cyanoxyl-mediated free radical polymerization (CMFRP) in a one-pot fashion. The aryl azide chain-end functionalized N-glycan polymers were characterized by 1H NMR and IR spectroscopy. The affinity-assisted photo-labeling capabilities of the aryl azide N-glycan polymers were demonstrated with aryl azide N-lactosyl polymer as a ligand for β-galactose-specific lectin from Arachis hypogaea (PNA) after UV irradiation and confirmed by SDS-PAGE with silver staining. Overall, the aryl azide chain-end functionalized N-linked glycan polymers will be useful multivalent ligands for specific protein labelling and functionality studies.

PubMed Disclaimer

Conflict of interest statement

There are no conflicts to declare.

Figures

Fig. 1
Fig. 1. Aryl azide chain-end functionalized glycopolymer for specific protein photo-labelling.
Scheme 1
Scheme 1. Straightforward synthesis of N-linked glycan polymers (4) from free glycans (1) via glycosylamine intermediates (2) followed by acrylation (3) and CMFRP.
Fig. 2
Fig. 2. 1H NMR spectrum of β-lactose (1), β-d-lactopyranosylamine (2), N-(prop-2-enoyl)-β-d-lactopyranosylamine (3), and p-azido-phenyl-β-d-Gal(1–4)-β-d-Glc-N-glycopolymer (4), (D2O, 400 MHz NMR).
Fig. 3
Fig. 3. FT-IR spectrum of p-Cl-phenyl-β-d-Gal(1–4)-β-d-Glc-N-glycopolymer (A) and p-azido-phenyl-β-d-Gal(1–4)-β-d-Glc-N-glycopolymer (B).
Fig. 4
Fig. 4. (A) SDS-PAGE characterization of glycopolymer–PNA crosslinking after UV irradiation. Lane M. MW marker; lane 1. Arachis hypogaea (PNA) only (negative control); lane 2. PNA + N3-AM polymer; lane 3. PNA + N3-Glc polymer; lane 4. PNA + N3-Gal polymer; lane 5. PNA + N3-Lac polymer. Silver staining, polymer/protein molar ratio: 6 : 1. (B) SDS-PAGE characterization of glycopolymer–ConA crosslinking after UV irradiation: lane M. MW marker; lane 1. ConA only (negative control); lane 2. ConA + N3-Glc polymer; lane 3. ConA + N3-Lac polymer. 50 ng of protein loaded in each lane. Silver staining.
Fig. 5
Fig. 5. (A) SDS-PAGE characterization of glycopolymer–protein crosslinking in controlled conditions after UV irradiation: lane M. MW marker; lane 1. N3-Lac polymer + free β-lactose + PNA (competitive); lane 2. PNA + free β-lactose + N3-Lac polymer (sequential); lane 3. N3-Lac polymer + PNA (1 : 1 molar ratio); lane 4. N3-Lac polymer + PNA (3 : 1 molar ratio); lane 5. N3-Lac polymer + PNA (6 : 1 molar ratio). (B) SDS-PAGE characterization of comparative glycopolymer–protein crosslinking after UV irradiation: lane M. MW marker lane 1. Cl-Lac polymer + PNA; lane 2. N3-Lac polymer + PNA. Silver staining. Polymer/protein molar ratio: 6 : 1.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Varki A. Glycobiology. 2017;27:3–49. doi: 10.1093/glycob/cww086. - DOI - PMC - PubMed
    1. Kiessling L. L. Splain R. A. Annu. Rev. Biochem. 2010;79:619–653. doi: 10.1146/annurev.biochem.77.070606.100917. - DOI - PubMed
    1. Wu H. Kohler J. Curr. Opin. Chem. Biol. 2019;53:173–182. doi: 10.1016/j.cbpa.2019.09.002. - DOI - PMC - PubMed
    1. Fujita A. Kohler J. J. Trends Glycosci. Glycotechnol. 2015;156:E1–E7.
    1. Wibowo A. Peters E. C. Hsieh-Wilson L. C. J. Am. Chem. Soc. 2014;136:9528–9531. doi: 10.1021/ja502482a. - DOI - PMC - PubMed