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Rapid and simple approach for the NMR resonance assignment of the carbohydrate chains of an intact glycoprotein Application of gradient‐enhanced natural abundance 1 H‐ 13 C HSQC and HSQC‐TOCSY to the α‐subunit of human chorionic gonadotropin
Author(s) -
de Beer Tonny,
van Zuylen Carol W.E.M.,
Hård Karl,
Boelens Rolf,
Kaptein Robert,
Kamerling Johannis P.,
Vliegenthart Johannes F.G.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00547-8
Subject(s) - heteronuclear single quantum coherence spectroscopy , anomer , chemistry , carbohydrate , monosaccharide , nmr spectra database , nuclear magnetic resonance spectroscopy , stereochemistry , spectral line , nuclear magnetic resonance , biochemistry , physics , astronomy
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 1 H‐ 13 C HSQC and HSQC‐TOCSY spectra to be recorded of the α‐subunit of human chorionic gonadotropin. Most carbohydrate 1 H‐ 13 C 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 1 H‐ 13 C correlations. Subsequently, correlations of the monosaccharide skeleton atoms were readily assigned in the HSQC‐TOCSY spectrum.