التفاصيل البيبلوغرافية
| العنوان: |
Total chemical synthesis and NMR characterization of the glycopeptide tx5a, a heavily post-translationally modified conotoxin, reveals that the glycan structure isα-d-Gal-(1→3)-α-d-GalNAc. |
| المؤلفون: |
Kang, James, Low, William, Norberg, Thomas, Meisenhelder, Jill, Hansson, Karin, Stenflo, Johan, Zhou, Guo-Ping, Imperial, Julita, Olivera, Baldomero M., Rigby, Alan C., Craig, A. Grey |
| المصدر: |
European Journal of Biochemistry; Dec2004, Vol. 271 Issue 23/24, p4939-4949, 11p |
| مصطلحات موضوعية: |
AMINO acids, ORGANIC acids, MASS spectrometry, MUSCLE diseases, CHROMATOGRAPHIC analysis, NUCLEAR magnetic resonance spectroscopy |
| مستخلص: |
The 13-amino acid glycopeptide tx5a (Gla-Cys-Cys-Gla-Asp-Gly-Trp*-Cys-Cys-Thr*-Ala-Ala-Hyp-OH, where Trp* = 6-bromotryptophan and Thr* = Gal-GalNAc-threonine), isolated fromConus textile, causes hyperactivity and spasticity when injected intracerebral ventricularly into mice. It contains nine post-translationally modified residues: four cysteine residues, twoγ-carboxyglutamic acid residues, and one residue each of 6-bromotryptophan, 4-trans-hydroxyproline and glycosylated threonine. The chemical nature of each of these has been determined with the exception of the glycan linkage pattern on threonine and the stereochemistry of the 6-bromotryptophan residue. Previous investigations have demonstrated that tx5a contains a disaccharide composed ofN-acetylgalactosamine (GalNAc) and galactose (Gal), but the interresidue linkage was not characterized. We hypothesized that tx5a contained the T-antigen,β-d-Gal-(1→3)-α-d-GalNAc, one of the most common O-linked glycan structures, identified previously in anotherConusglycopeptide, contalukin-G. We therefore utilized the peracetylated form of this glycan attached to Fmoc-threonine in an attempted synthesis. While the result-ing synthetic peptide (Gla-Cys-Cys-Gla-Asp-Gly-Trp*-Cys-Cys-Thr*-Ala-Ala-Hyp-OH, where Trp* =6-bromotryptophan and Thr* = β-d-Gal-(1→3)-α-d-GalNAc-threonine) and the native peptide had almost identical mass spectra, a comparison of their RP-HPLC chromatograms suggested that the two forms were not identical. Two-dimensional1H homonuclear and13C-1H heteronuclear NMR spectroscopy of native tx5a isolated fromConus textilewas then used to determine that the glycan present on tx5a indeed is not the aforementioned T-antigen, but ratherα-d-Gal-(1→3)-α-d-GalNAc. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
