Stable isotope labeling approaches for NMR characterization of glycoproteins using eukaryotic expression systems

Journal of Biomolecular NMR
Saeko YanakaKoichi Kato

Abstract

Glycoproteins are characterized by the heterogeneous and dynamic nature of their glycan moieties, which hamper crystallographic analysis. NMR spectroscopy provides potential advantages in dealing with such complicated systems, given that the target molecules can be isotopically labeled. Methods of metabolic isotope labeling in recombinant glycoproteins have been developed recently using a variety of eukaryotic production vehicles, including mammalian, yeast, insect, and plant cells, each of which has a distinct N-glycan diversification pathway. Yeast genetic engineering has enabled the overexpression of homogeneous high-mannose-type oligosaccharides with 13C labeling for NMR characterization of their conformational dynamics. The utility of stable isotope-assisted NMR spectroscopy has also been demonstrated using the Fc fragment of immunoglobulin G (IgG) as a model glycoprotein, providing useful information regarding intramolecular carbohydrate-protein interactions. Transverse relaxation optimization of intact IgG with a molecular mass of 150 kDa has been achieved by tailored deuteration of selected amino acid residues using a mammalian expression system. This offers a useful probe for the characterization of molecular interacti...Continue Reading

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Citations

May 16, 2020·Biophysics Reviews·Saeko YanakaKoichi Kato
Jul 26, 2018·Journal of Biomolecular NMR·Jerome Boisbouvier, Lewis E Kay
Dec 10, 2019·MedChemComm·Pablo ValverdeAna Gimeno
Feb 5, 2019·Frontiers in Molecular Biosciences·Steffen P Graether
Dec 15, 2020·Nature Communications·Alistair M JaggerDavid A Lomas
Dec 31, 2020·International Journal of Molecular Sciences·Yuji Tokunaga, Koh Takeuchi
Sep 5, 2020·Progress in Nuclear Magnetic Resonance Spectroscopy·Iva PritišanacPeter Güntert
Mar 22, 2021·The Journal of Biological Chemistry·James H Prestegard
Nov 1, 2020·Current Opinion in Structural Biology·Luca UnioneOscar Millet
Oct 2, 2019·ACS Central Science·Luca UnioneOscar Millet
Sep 10, 2019·ACS Omega·Pablo ValverdeJesús Jiménez-Barbero
May 18, 2020·Biochimica Et Biophysica Acta. Biomembranes·Gaddafi I Danmaliki, Peter M Hwang

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