Preparation and Structural Analysis of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell (RBC) Membranes
Main Article Content
Abstract
Glycophorins (GPs) in the red blood cell (RBC) membranes of carp (Cyprinus carpio L.) exhibit bacteriostatic activity against various gram-negative and gram-positive bacteria. This antibacterial property is attributed to a sialo-oligosaccharide. For a structural analysis of this O-linked sialo-oligosaccharide, it is necessary to prepare and purify the oligosaccharide from carp GPs. To prepare sialo-oligosaccharides, RBC membrane proteins were solubilized, and anion exchange column chromatography was performed to obtain the GPs. The GP yield from carp RBC membranes was ca.100% as total sialic acid. Then, released oligosaccharides were obtained from GP preparation by β-elimination. Second, the resulting oligosaccharide fraction was separated into two sialo-oligosaccharide components (P-1 and P-2) using the same anion exchange column. The oligosaccharide preparations that contained high amounts of NaCl were desalted with an activated charcoal column combined with ammonium bicarbonate in acetonitrile as an eluent. Using NMR to analyse the structure of the antimicrobial P-1, we determined the alignment of each proton and C1 carbon. To determine the linkage sites, we performed GC-MS using the permethylated P-1. Lastly, we determined that the structure of P-1 was NeuGcα2→6 (Fucα1→4) (Glcα1→3) Galβ1→4GalNAc-ol.
Keywords:
carp, glycophorin, sialo-oligosaccharide, column chromatography, NMR, GC‒MS
Article Details
How to Cite
AOKI, Takahiko.
Preparation and Structural Analysis of a Sialo-Oligosaccharide from Glycophorin in Carp Red Blood Cell (RBC) Membranes.
Medical Research Archives, [S.l.], v. 11, n. 3, mar. 2023.
ISSN 2375-1924.
Available at: <https://esmed.org/MRA/mra/article/view/3644>. Date accessed: 04 dec. 2024.
doi: https://doi.org/10.18103/mra.v11i3.3644.
Section
Research Articles
The Medical Research Archives grants authors the right to publish and reproduce the unrevised contribution in whole or in part at any time and in any form for any scholarly non-commercial purpose with the condition that all publications of the contribution include a full citation to the journal as published by the Medical Research Archives.
References
1. Cartron J-P, Colin Y, Kudo S, Fukuda M. Molecular genetics of Human Erythrocyte Sialoglycoproteins Glycophorins A, B, C, and D. In Blood Cell Biochemistry vol. 1, 1990
(pp.299-335). Springer USA.
2. Blanchard D. Human red cell glycophorins: Biochemical and antigenic properties. Transfusion Medicine Reviews. 1990;4:170-186.
3. Chasis JA, Mohandas N. Red blood cell glycophorins. Blood. 1992;80:1869-1879.
4. Tanner MJA. Molecular and cellular biology of the erythrocyte anion exchanger (AE1). Seminars in Hematology. 1993;30:34-57.
5. Yamashita T, Murayama J, Utsumi H, Hamada A. Structural studies of O-glycosidic oligosaccharide units of dog erythrocyte glycophorin. Biochim. Biophys. Acta. 1985;839:26-31.
6. Angel A-S, Grönberg G, Krotkiewski H, Lisowska E, Nilsson B. Structural analysis of the N-linked oligosaccharides from murine glycophorin. Arch. Biochem. Biophys. 1991; 291:76-88.
7. Murayama J-I, Utsumi H, Hamada A. Amino acid sequence of monkey erythrocyte glycophorin MK. Its amino acid sequence has a striking homology with that of human glycophorin A. Biochim. Biophys. Acta. 1989;999:273-280.
8. Krotkiewski H, The structure of glycophorins of animal erythrocytes. Glycoconj. J. 1988;5;35-48.
9. Watts C, Wheeler KP. Partial separation of a sodium-dependent transport system for amino acids in avian erythrocyte membranes. FEBS Letters. 1978;94:241-244.
10. Dockham PA, Vidaver GA. Comparison of human and pigeon erythrocyte membrane proteins by one and two-dimensional gel electrophoresis. Comparative Biochemistry and Physiology Part B. 1987;87:171-177.
11. Aoki T, Chimura K, Nakao N, Mizuno Y. Isolation and characterization of glycophorin from carp red blood cell membranes. Membranes. 2014;4:491-508.
12. 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.
13. Aoki T. Determination of the bacteriostatic activity of glycophorin preparations from red blood cell (RBC) membranes of yellow tail and red sea bream. Medical Research Archives.
2021;9. doi: 10.18103/mra.v9i12.2613.
14. Aoki T. Behaviour of a sialo-oligosaccharide from glycophorin in teleost red blood cell membranes. In Animal models and experimental research in medicine, 2022; doi: 10.5772/intechopen.107234.
15. Michel F, Rudloff V. Isolation and characterization of the rainbow trout erythrocyte band-3 protein. Eur. J. Biochem. 1989;181:181-187.
16. Marchesi VT, Andrews EP. Glycoproteins: Isolation from cell membranes with lithium
diiodosalicylate. Science. 1971;174:1247-1248.
17. Honma K, Manabe H, Tomita M, Hamada A. Isolation and preliminary characterization of two glycophorins from rabbit erythrocyte membranes. Chem. Pharm. Bull. 1982;30:966-972.
18. Aoki T. Preparation method of carp origin antibacterial agent and antibacterial device using this agent (In Japanese). Japanese patent (Patent application: 2009-95745).
19. Carlson DM. Structures and immunochemical properties of oligosaccharides isolated from pig submaxillary mucins. J. Biol. Chem. 1968;243:616–626.
20. Morelle W, Strecker G. Structural analysis of oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Bufo bufo: characterization of the carbohydrate sequence Gal(α1-3)GalNAc(α1-3)[Fuc(α1-2)]Gal. Glycobiology. 1997;7:777-790.
21. Hokke CH, Bergwerff AA, Van Dedem GWK, Kamerling JP, Vliegenthart JFG. Structural analysis of the sialylated N- and O-linked carbohydrate chains of recombinant human erythropoietin expressed in Chinese hamster ovary cells: Sialylation patterns and branch location of dimeric N-acetyllactosamine units. Eur. J. Biochem. 1995;228:981-1008.
22. Kabir S, Gerwig GJ. The structural analysis of the O-glycans of the jacalin-bound rabbit immunogloblin G. In Biochemistry and molecular biology international, Vol 42, 1997(pp769-778). Academic Press, Inc. Australia.
23. Morelle W, Guyétant R, Strecker G. Structural analysis of oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Rana dalmatina. Carbohydrate Research. 1998;306:435-443.
24. Lindstad RI, Köll P, Mckinley-Mckee JS. Substrate specificity of sheep liver sorbitol dehydrogenase. Biochem. J. 1998;330:479-487.
25. Packer NH, Lawson MA, Jardine DR, Redmond JW. A general approach to desalting
oligosaccharides released from glycoproteins. Glycoconj. J. 1998;15:737–747.
26. Ciucanu I, Kerek F. A simple and rapid method for the permethylation of carbohydrates. Carbohydr. Res. 1984;131:209–217.
27. Vliegenthart JFG, Dorland L, van Halbeek H. High-resolution, 1H-nuclear magnetic resonance spectroscopy as a tool in the structural analysis of carbohydrates related to glycoproteins. In Advances in carbohydrate chemistry and biochemistry, Vol 41, 1983 (pp.209-373). Academic Press, Inc. USA.
28. Imberty A, Oligosaccharide structure: theory versus experiment. Current Opinion in Stuructural Biology. 1997;7:617-623.
29. Fukuda M, Lauffenburger M, Sasaki H, Rogers ME, Dell A. Structures of novel sialylated O-linked oligosaccharides isolated from human erythrocyte glycophorins. J. Biol. Chem. 1987;262:11952–11957.
30. Duk M, Krotkiewski H, Stasyk TV, Lutsik-Kordovsky M, Syper D, Lisowska E. Isolation and characterization of glycophorin from nucleated (Chicken) erythrocytes. Arch. Biochem. Biophys. 2000;375:111–118.
31. Guérardel Y, Balanzino L, Maes E, Leroy Y, Coddeville B, Oriol R, Strecker G. The nematode Caenorhabditis elegans synthesizes unusial O-linked glycans: Identification of glucose-substituted mucin-type O-glycans and short chondroitin-like oligosaccharides. Biochem. J. 2001;357:167–182.
32. Podbielska M, Fredriksson S-Å, Nilsson B, Lisowska E, Krotkiewski H. ABH blood group antigens in O-glycans of human glycophorin A. Arch. Biochem. Biophys. 2004;429:145–153.
33. Araki T, Aoki T, Kitamikado M. Isolation and Identification of a β-1,3-xylanase-producing bacterium. Nippon Suisan Gakkaishi 1987;53:2077–2081.
34. Prohaska R, Koerner TAW Jr, Armitage IM, Furthmayr H. Chemical and carbon-13 nuclear magnetic resonance studies of the blood group M and N active sialoglycopeptides from human glycophorin A. J. Biol. Chem. 1981;256:5781–5791.
35. Aoki T. A comprehensive review of our current understanding of red blood cell (RBC) glycoproteins. Membranes 2017;7:56–74
(pp.299-335). Springer USA.
2. Blanchard D. Human red cell glycophorins: Biochemical and antigenic properties. Transfusion Medicine Reviews. 1990;4:170-186.
3. Chasis JA, Mohandas N. Red blood cell glycophorins. Blood. 1992;80:1869-1879.
4. Tanner MJA. Molecular and cellular biology of the erythrocyte anion exchanger (AE1). Seminars in Hematology. 1993;30:34-57.
5. Yamashita T, Murayama J, Utsumi H, Hamada A. Structural studies of O-glycosidic oligosaccharide units of dog erythrocyte glycophorin. Biochim. Biophys. Acta. 1985;839:26-31.
6. Angel A-S, Grönberg G, Krotkiewski H, Lisowska E, Nilsson B. Structural analysis of the N-linked oligosaccharides from murine glycophorin. Arch. Biochem. Biophys. 1991; 291:76-88.
7. Murayama J-I, Utsumi H, Hamada A. Amino acid sequence of monkey erythrocyte glycophorin MK. Its amino acid sequence has a striking homology with that of human glycophorin A. Biochim. Biophys. Acta. 1989;999:273-280.
8. Krotkiewski H, The structure of glycophorins of animal erythrocytes. Glycoconj. J. 1988;5;35-48.
9. Watts C, Wheeler KP. Partial separation of a sodium-dependent transport system for amino acids in avian erythrocyte membranes. FEBS Letters. 1978;94:241-244.
10. Dockham PA, Vidaver GA. Comparison of human and pigeon erythrocyte membrane proteins by one and two-dimensional gel electrophoresis. Comparative Biochemistry and Physiology Part B. 1987;87:171-177.
11. Aoki T, Chimura K, Nakao N, Mizuno Y. Isolation and characterization of glycophorin from carp red blood cell membranes. Membranes. 2014;4:491-508.
12. 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.
13. Aoki T. Determination of the bacteriostatic activity of glycophorin preparations from red blood cell (RBC) membranes of yellow tail and red sea bream. Medical Research Archives.
2021;9. doi: 10.18103/mra.v9i12.2613.
14. Aoki T. Behaviour of a sialo-oligosaccharide from glycophorin in teleost red blood cell membranes. In Animal models and experimental research in medicine, 2022; doi: 10.5772/intechopen.107234.
15. Michel F, Rudloff V. Isolation and characterization of the rainbow trout erythrocyte band-3 protein. Eur. J. Biochem. 1989;181:181-187.
16. Marchesi VT, Andrews EP. Glycoproteins: Isolation from cell membranes with lithium
diiodosalicylate. Science. 1971;174:1247-1248.
17. Honma K, Manabe H, Tomita M, Hamada A. Isolation and preliminary characterization of two glycophorins from rabbit erythrocyte membranes. Chem. Pharm. Bull. 1982;30:966-972.
18. Aoki T. Preparation method of carp origin antibacterial agent and antibacterial device using this agent (In Japanese). Japanese patent (Patent application: 2009-95745).
19. Carlson DM. Structures and immunochemical properties of oligosaccharides isolated from pig submaxillary mucins. J. Biol. Chem. 1968;243:616–626.
20. Morelle W, Strecker G. Structural analysis of oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Bufo bufo: characterization of the carbohydrate sequence Gal(α1-3)GalNAc(α1-3)[Fuc(α1-2)]Gal. Glycobiology. 1997;7:777-790.
21. Hokke CH, Bergwerff AA, Van Dedem GWK, Kamerling JP, Vliegenthart JFG. Structural analysis of the sialylated N- and O-linked carbohydrate chains of recombinant human erythropoietin expressed in Chinese hamster ovary cells: Sialylation patterns and branch location of dimeric N-acetyllactosamine units. Eur. J. Biochem. 1995;228:981-1008.
22. Kabir S, Gerwig GJ. The structural analysis of the O-glycans of the jacalin-bound rabbit immunogloblin G. In Biochemistry and molecular biology international, Vol 42, 1997(pp769-778). Academic Press, Inc. Australia.
23. Morelle W, Guyétant R, Strecker G. Structural analysis of oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Rana dalmatina. Carbohydrate Research. 1998;306:435-443.
24. Lindstad RI, Köll P, Mckinley-Mckee JS. Substrate specificity of sheep liver sorbitol dehydrogenase. Biochem. J. 1998;330:479-487.
25. Packer NH, Lawson MA, Jardine DR, Redmond JW. A general approach to desalting
oligosaccharides released from glycoproteins. Glycoconj. J. 1998;15:737–747.
26. Ciucanu I, Kerek F. A simple and rapid method for the permethylation of carbohydrates. Carbohydr. Res. 1984;131:209–217.
27. Vliegenthart JFG, Dorland L, van Halbeek H. High-resolution, 1H-nuclear magnetic resonance spectroscopy as a tool in the structural analysis of carbohydrates related to glycoproteins. In Advances in carbohydrate chemistry and biochemistry, Vol 41, 1983 (pp.209-373). Academic Press, Inc. USA.
28. Imberty A, Oligosaccharide structure: theory versus experiment. Current Opinion in Stuructural Biology. 1997;7:617-623.
29. Fukuda M, Lauffenburger M, Sasaki H, Rogers ME, Dell A. Structures of novel sialylated O-linked oligosaccharides isolated from human erythrocyte glycophorins. J. Biol. Chem. 1987;262:11952–11957.
30. Duk M, Krotkiewski H, Stasyk TV, Lutsik-Kordovsky M, Syper D, Lisowska E. Isolation and characterization of glycophorin from nucleated (Chicken) erythrocytes. Arch. Biochem. Biophys. 2000;375:111–118.
31. Guérardel Y, Balanzino L, Maes E, Leroy Y, Coddeville B, Oriol R, Strecker G. The nematode Caenorhabditis elegans synthesizes unusial O-linked glycans: Identification of glucose-substituted mucin-type O-glycans and short chondroitin-like oligosaccharides. Biochem. J. 2001;357:167–182.
32. Podbielska M, Fredriksson S-Å, Nilsson B, Lisowska E, Krotkiewski H. ABH blood group antigens in O-glycans of human glycophorin A. Arch. Biochem. Biophys. 2004;429:145–153.
33. Araki T, Aoki T, Kitamikado M. Isolation and Identification of a β-1,3-xylanase-producing bacterium. Nippon Suisan Gakkaishi 1987;53:2077–2081.
34. Prohaska R, Koerner TAW Jr, Armitage IM, Furthmayr H. Chemical and carbon-13 nuclear magnetic resonance studies of the blood group M and N active sialoglycopeptides from human glycophorin A. J. Biol. Chem. 1981;256:5781–5791.
35. Aoki T. A comprehensive review of our current understanding of red blood cell (RBC) glycoproteins. Membranes 2017;7:56–74