1H and 13C NMR assignments for the glycans in glycoproteins by using 2H/13C-labeled glucose as a metabolic precursor.
In order to understand the role of the glycans in glycoproteins in solution, structural information obtained by NMR spectroscopy is obviously required. However, the assignment of the NMR signals from the glycans in larger glycoproteins is still difficult, mainly due to the lack of appropriate methods for the assignment of the resonances originating from the glycans. By using [U-13C6, 2H7]glucose as a metabolic precursor, we have successfully prepared a glycoprotein whose glycan is uniformly labeled with 13C and partially with D at the sugar residues. The D to H exchange ratios at the C1-C6 positions of the sugar residues have been proven to provide useful information for the spectral assignments of the glycan in the glycoprotein. This is the first report on the residue-specific assignment of the anomeric resonances originating from a glycan attached to a glycoprotein by using the metabolic incorporation of hydrogen from the medium into a glycan labeled with [U-13C6, 2H7]glucose.
Yamaguchi
Y
Y
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan.
Takizawa
T
T
Kato
K
K
Arata
Y
Y
Shimada
I
I
D000818:Animals; D002247:Carbon Isotopes; D003903:Deuterium / Q000378:metabolism; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D011522:Protons
D002247:Carbon Isotopes; D006023:Glycoproteins; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D011134:Polysaccharides; D011522:Protons; D003903:Deuterium; D005947:Glucose