Structure of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell Membranes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Preparation and Isolation of Glycophorin from the Red Blood Cell Membranes
2.3. Preparation of the Carp Glycophorin Carbohydrate Fraction
2.4. High-Performance Liquid Chromatography (HPLC)
2.5. Desalting of Oligosaccharide Fractions
2.6. Thin-Layer Chromatography (TLC)
2.7. Neutral Sugar Analysis
2.8. NMR and GC-MS
3. Results
3.1. Homogeneity of P-1 and P-2 Fractions
3.2. Neutral Sugar Composition
3.3. NMR and GC-MS
Residue | Reporter Site | 1H | 13C |
---|---|---|---|
GalNAc-ol | 1a 1e | 3.626 3.763 | 67.34 67.34 |
2 | 4.130 | 69.21 | |
3 | 3.592 | 67.58 | |
4 | 3.749 | 53.66 | |
5 | 3.519 | 71.71 | |
6 | 3.326 | 37.82 | |
αGlc | 1 | 5.103 | 91.01 |
2 | 4.012 | 75.42 | |
3 | 3.905 | 82.62 | |
4 | 3.567 | 75.20 | |
5 | 3.727 | 70.37 | |
6 | 3.803 | 67.32 | |
αFuc | 1 | 4.917 | 106.5 |
2 | 4.048 | 62.64 | |
3 | 3.875 | 68.61 | |
4 | 3.678 | 61.08 | |
5 | 4.764 | 71.60 | |
-CH3 | 1.234 | 20.10 | |
βGal | 1 | 4.518 | 95.39 |
2 | 3.726 | 73.70 | |
3 | 3.583 | 62.76 | |
4 | 3.583 | 69.60 | |
5 | 4.007 | 50.27 | |
6 | 4.023 | 70.04 |
Peak No. | Linkage Site | GC Retention Time (min) | Peak Area | MS Retention Time (min) |
---|---|---|---|---|
1 | − | 15.297 | 13,928 | 13.23 |
2 | Pen 1 → | 18.747 | 11,924 | 16.47 |
3 | → 2 Pen 1 → | 20.200 | 7,818 | 18.13 |
4 | Fuc 1 → | 20.747 | 13,925 | 18.42 |
5 | → 5 Pen 1 → | 21.170 | 30,312 | 19.09 |
6 | Hex 1 → | 21.755 | 26,944 | 19.43 |
7 | Hex 1 → | 22.071 | 41,924 | 20.02 |
8 | → 6 Hex 1 → | 22.153 | 28,993 | 20.04 |
9 | → 6 Hex 1 → | 22.205 | 8,515 | 20.07 |
10 | → 4 Hex 1 → | 23.162 | 91,934 | 21.05 |
11 | → 4 Hex 1 → | 23.257 | 39,668 | 21.10 |
12 | → 3 Hex 1 → | 23.361 | 25,210 | 21.17 |
13 | → 3,4 Hex 1 → | 24.083 | 39,414 | 21.59 |
14 | → 4 HexNAc 1 → | 27.002 | 11,999 | 24.43 |
4. Discussion
Abbreviations
NeuGc | N-glycolylneuraminic acid |
Ac | acetyl |
Fuc | fucose |
Glc | glucose |
Gal | galactose |
GalNAc-ol | N-acetylgalactosaminitol |
Acknowledgments
Conflicts of Interest
References
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Aoki, T.; Chimura, K.; Sugiura, H.; Mizuno, Y. Structure of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell Membranes. Membranes 2014, 4, 764-777. https://doi.org/10.3390/membranes4040764
Aoki T, Chimura K, Sugiura H, Mizuno Y. Structure of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell Membranes. Membranes. 2014; 4(4):764-777. https://doi.org/10.3390/membranes4040764
Chicago/Turabian StyleAoki, Takahiko, Kenji Chimura, Hikaru Sugiura, and Yasuko Mizuno. 2014. "Structure of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell Membranes" Membranes 4, no. 4: 764-777. https://doi.org/10.3390/membranes4040764