NMR Spectroscopy of Sialic Acids

Vliegenthart, Johannes F.G.; Dorland, L.; Halbeek, Herman van; Haverkamp, Johan

doi:10.1007/978-3-7091-8680-0_7

Cited by 178 publications

(268 citation statements)

References 49 publications

Supporting

Mentioning

241

Contrasting

Unclassified

Order By: Relevance

“…These assignments were based upon the observation of the C3H2.C4H.C5H spin system in DQF-COSY and TOCSY spectra and upon appropriate connectivities to an amide proton signal in DQF-COSY, TOCSY, and NOESY spectra. The chemical shifts of the signals are all within 0.2 ppm of the expected values for sialic acid in an a-glycosidic linkage to Gal (Vliegenthart et al, 1981). The extremely sharp lines observed for all the sialic acid signals is consistent with the presumed chain-terminal position of these residues and their consequent expected high mobility.…”

Section: Carbohydrate Assignmentssupporting

confidence: 68%

Sequence‐specific

Fletcher¹,

Harrison²,

Lachmann³

et al. 1993

Protein Science

View full text Add to dashboard Cite

CD59 is a recently discovered cell-surface glycoprotein that restricts lysis by homologous complement and has limited sequence similarity to snake venom neurotoxins. This paper describes the first results of a two-dimensional NMR study of CD59 prepared from human urine. Nearly complete 'H-NMR assignments were obtained for the 77 amino acid residues and partial assignments for the N-glycan and the glycosylphosphatidylinositol (GPI) anchor. These results together confirm that the C-terminal residue of the mature protein is Asn 77 and that the urine-derived form retains the nonlipid part of the GPI anchor. The data further indicate that the GPI anchor and possibly the N-glycan are structurally inhomogeneous and suggest that the phospholipid present in the intact GPI anchor was removed by phosphatidylinositol-specific phospholipase-D. The folding topology of the protein was determined from NOE enhancements and slowly exchanging backbone amide protons and consists primarily of five extended strands (denoted p l -f l 5 in sequence order), arranged into separate two-stranded (PI and &) and three-stranded (&&) antiparallel P-sheets. The same folding topology is found in all of the snake venom neurotoxins whose structures have been determined. The region between the & and & strands has helical character, a feature that is not present in the neurotoxins but that is seen in the topologically similar wheat germ agglutinin.

show abstract

Section: Carbohydrate Assignmentssupporting

confidence: 68%

Sequence‐specific

Fletcher¹,

Harrison²,

Lachmann³

et al. 1993

Protein Science

View full text Add to dashboard Cite

show abstract

“…'H chemical shifts are given at 300 K by reference to intemal acetone (6 2.225; Vliegenthart et al, 1983), while 13C chemical shifts are given by reference to the a-anomeric resonance of I3C-labeled glucose (6 92.9; Bock & Pedersen, 1983). The 'H chemical shifts and the vicinal 'H-'H coupling constants were determined from 600-MHz 1D spectra, except for H5 and H6 whose 'H chemical shifts were determined from the 2D TOCSY spectrum.…”

Section: H6"mentioning

confidence: 99%

“…The 13C chemical shifts were determined from the 'H-13C correlations in the 2D IH-I3C HSQC spectrum. The notation for tryptophan is as described by Wiithrich (1986), and for the hexose moiety as described by Vliegenthart et al (1983). Might be interchanged with the 13C6 chemical shift of W-10 (6 122.6).…”

Section: H6"mentioning

confidence: 99%

New type of linkage between a carbohydrate and a protein: C-glycosylation of a specific tryptophan residue in human RNase Us

Hofsteenge¹,

Müller²,

Beer³

et al. 1994

Biochemistry

Self Cite

263

234

View full text Add to dashboard Cite

We report a new type of linkage between a carbohydrate and a protein, involving the rarely modified side chain of a tryptophan residue. An aldohexopyranosyl residue was found to be linked via a C-C bond to the indole ring of the tryptophan residue at position 7 of human RNase Us. Mass spectrometric analysis of peptides containing this residue showed a molecular mass 162 Da higher than that expected for tryptophan. The fragmentation pattern of the modified amino acid side chain was reminiscent of that of aromatic C-glycosides, suggesting a direct attachment of a hexose residue to a C-position of the tryptophan indole moiety. 'H and 13C NMR spectroscopic data confirmed this inference and unequivocally demonstrated the substituent to be an aldohexopyranosyl residue, C-glycosidically linked to the C2 atom of the indole. This mode of attachment differs from the ones known so far, in which carbohydrates are linked to an amino acid side chain by N-or 0-glycosidic bonds.

show abstract

“…cChromatographrc analyses for GP2 (i) and GPZ2 (11). Those under (III) are calculated from the n.m.r.…”

Section: Carbohydrate and Amino Acid Composition -mentioning

confidence: 99%

Structural analysis of the carbohydrate moieties of human tamm-horsfall glycoprotein

Williams

Marshall

Halbeek

et al. 1984

Carbohydrate Research

Self Cite

View full text Add to dashboard Cite

ABSTRAaGlycopeptides present in a pronase digest of human Tamm-Horsfall glycoprotein were fractionated by chromatography on DEAE-Sephadex A25 in 0.1~ acetic acid. The separated glycopeptides were characterised by 500-MHz 'H-n.m.r. spectroscopy, in conjunction with sugar and amino acid analysis, and they were shown to be of the N-glycosylic, N-acetyllactosamine type. Each fraction consisted mainly of a tetra-antennary entity having various degrees of complexity, with lesser amounts of the triantennary structure, and even smaller amounts of the diantennary type. There was extensive heterogeneity in non-reducing terminal groups in each of the glycopeptides, whereas the peptide portions were similar. The extent to which any one of the galactose residues in the N-acetyllactosamine units was substituted, and the type of substitution, differed. The substituents were cr-NeuAc-(2-+6) 9 a-NeuAc-(2+3), and a-NeuAc-(2+3) ]. The carbohydrate moieties of the glycoprotein were heterogeneous also because of an uneven distribution of the fucose residues, which were attached to GlcNAc residues, both that linked to asparagine and one or more of those present in the N-acetyllactosamine units. The cr-NeuAc-(2+3)[@GalNAc-(1+4)]-P-Gal-(l+ sequence forms, at least in part, the Sd" immunodeterminant.The pK, of the carboxyl group of the sialic acid residue in this entity is lower than that for molecules lacking GalNAc in this position. Thus, the difference in the number of Sd" determinants carried by the glycopeptides enabled the latter to be fractionated on DEAESephadex.

show abstract

NMR Spectroscopy of Sialic Acids

Cited by 178 publications

References 49 publications

Sequence‐specific

Sequence‐specific

New type of linkage between a carbohydrate and a protein: C-glycosylation of a specific tryptophan residue in human RNase Us

Structural analysis of the carbohydrate moieties of human tamm-horsfall glycoprotein

Contact Info

Product

Resources

About