Rapid and simple approach for the NMR resonance assignment of the carbohydrate chains of an intact glycoprotein Application of gradient‐enhanced natural abundance 1H‐13C HSQC and HSQC‐TOCSY to the α‐subunit of human chorionic gonadotropin

FEBS Letters

Boelens, Rolf¹  Hård, Karl¹  Kaptein, Robert¹  de Beer, Tonny²  Vliegenthart, Johannes F.G.²  Kamerling, Johannis P.²  van Zuylen, Carol W.E.M.² 

[1]NMR Spectroscopy, Utrecht University, Padualaan 8, NL-3584 CH Utrecht, The Netherlands

[2]Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Padualaan 8, NL-3584 CH Utrecht, The Netherlands

关键词: Glycoprotein;    α-subunit;    Human chorionic gonadotropin;    Pulsed field gradients;    Nuclear magnetic resonance;    1H-13C correlation spectroscopy;    NMR;    nuclear magnetic resonance;    HSQC;    1H-detected heteronuclear single quantum coherence spectroscopy;    TOCSY;    total correlation spectroscopy;    INEPT;    insensitive nuclei enhanced by polarization transfer;    MLEV;    M. Levitt;    GARP;    globally optimized alternating-phase rectangular pulses;    hCG;    human chorionic gonadotropin;    Gal;    D-galactose;    Man;    D-mannose;    GIcNAc;    N-acetyl-D-glucosamine;    NeuAc;    N-acetyl-D-neuraminic acid;   

DOI  :  10.1016/0014-5793(94)00547-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The structure assessment of an intact glycoprotein in solution requires an extensive assignment of the carbohydrate NMR resonances. However, assignment of homonuelear spectra is very complicated because of the severe overlap of protein and carbohydrate signals. Application of pulsed field gradients allowed high quality natural abundance ¹H-¹³C HSQC and HSQC-TOCSY spectra to be recorded of the α-subunit of human chorionic gonadotropin. Most carbohydrate ¹H-¹³C correlations appear in a distinct region between the aromatic region and the protein C^α-H^α region. The enormous reduction in overlap led to fast and unambiguous assignment of the anomeric ¹H-¹³C correlations. Subsequently, correlations of the monosaccharide skeleton atoms were readily assigned in the HSQC-TOCSY spectrum.

【 授权许可】

Unknown