The potency of amide protons for assignments of NMR spectra of carbohydrate chains of glycoproteins, recorded in 1H2O solutions.
Three glycoprotein N-glycans, namely, a disialylated diantennary carbohydrate chain linked to Asn, a monosialylated, fucosylated diantennary glycopeptide with bisecting N-acetylglucosamine, and a tetrasialylated, fucosylated tetra-antennary oligosaccharide, have been investigated by two-dimensional NOE and HOHAHA spectroscopy in 1H2O as solvent. The amide protons of all N-acetylglucosamine and sialic acid residues could readily be assigned. The large chemical-shift dispersion of the amide resonances of the N-acetylglucosamine residues, allowed the unambiguous assignment of every N-acetyl methyl signal, via strong NOEs. Subspectra could be obtained of all N-acetylglucosamine residues in HOHAHA spectra. These results have as main implication that several biologically important large glycans will now [corrected] become amenable for conformational studies by multidimensional NMR in 1H2O solution.
Hård
K
K
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
Spronk
B A
BA
Hokke
C H
CH
Kamerling
J P
JP
Vliegenthart
J F
JF
D000117:Acetylglucosamine / Q000737:chemistry; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D004187:Disaccharides; D005936:Glucans / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid; D011134:Polysaccharides / Q000737:chemistry; D012794:Sialic Acids / Q000737:chemistry; D012996:Solutions; D014867:Water
D002241:Carbohydrates; D004187:Disaccharides; D005936:Glucans; D006023:Glycoproteins; D011134:Polysaccharides; D012794:Sialic Acids; D012996:Solutions; D014867:Water; C010913:N,N-diacetylchitobiose; D019158:N-Acetylneuraminic Acid; D000117:Acetylglucosamine