1995 Fiscal Year Final Research Report Summary
LOCALIZATION OF ABO-TYPE GLYCOSYLTRANSFERASES IN CELLS AND SUBSTRATE DISTRUBUTION
| Project/Area Number |
06670161
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
KUSHI Yasunori Tokyo Medical and Dental University, Faculty of Medicine, Reserach Associate, 医学部, 助手 (60177988)
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| Co-Investigator(Kenkyū-buntansha) |
ROKUKAWA Chiaki Tokyo Medical and Dental University, Faculty of Medicine, Technical assistant, 医学部, 技官
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| Project Period (FY) |
1994 – 1995
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| Keywords | ABO-type blood group, / glycosyltransferase, / al-3, N-acetylgalactosamine / blood group active glycosphingolipid, / ganglioside |
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| Research Abstract |
Total neutral glycolipids and gangliosides in Folch's upper phase were isolated from human erythrocyte membranes of 22 individuals with blood group type AB.On immunostaining after TLC with an anti-A antibody, all reactive glycolipids in type A erythrocytes were found to correspond to reactive glycolipids in type AB erythrocytes. With an anti-B antibody, all reactive glycolipids in type B erythrocytes were also found to correspond to reactive glycolipids in type AB erythrocytes. On comparison of type AB erythrocytes reactivity to anti-A and anti-B antibodies, it was found that all glycolipids reactive with either the anti-A or the anti-B antibody were detected in type A and type B erythrocytes. Also, on TLC A-active gangliosides exhibited R_f values a little higher than those of B-active gangliosides.
A series of gangliosides reactive with antibodies was separated from Folch's upper gangliosides by DEAE-Sephadex ion exchange chromatography and characterized by TLC immunostaining. The results obtained strongly suggested that A-and B-active carbohydrate chain epitopes were present as separate ganglioside molecules in type AB erythrocytes. Furthermore, upon immunoprecipitation treatment of fractions obtained on column chromatography against the anti-A antibody, branched hybrid-type molecules with both A-and B-determinants were not detected.
The existence of blood group antigens in both the neutral glycolipid and ganglioside fractions strongly indicates that the antigenicity of the blood group antigen in AB type erythrocytes was due not only to neutral glycolipids but also influenced by gangliosides, and that the fine regulation of the erythrocyte surface is possibly controlled by the degree of sialylation of blood group active glycosphingolipids. The novel A-active gangliosides were purified from type A erythrocytes membranes by using conventional methods. Now those detailed structures and functions are anakyzed.
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[Publications] Ariga,T.,Tao,R.V.,Lee,B.-C.,Yamawaki,M.,Yoshino,H.,Scarsdale,N.J.,Kasama,T.,Kushi,Y.and Yu,R.K.: "Glycolipid Composition of Human Cataractous Lenses:Characterization of Lewisx glycolipids" J.Biol.Chem.269. 2667-2675 (1994)
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[Publications] Watarai,H.,Kushi,Y.,Shigeto,R.,Misawa,N.,Eishi,Y.,Handa,S.,and Yasuda,T.: "Production of Monoclonal Antibody Directed to Hanganutziu-Deichger Active Gangliosides,N-glycolylneuraminic Acid-Containing Gangliosides" J.Biochem.(Tokyo). 117. 1062-1069 (1995)
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[Publications] Kushi,Y.,Tsunoda,A.,Komatsuzaki,A.,Watanabe,K.,Kasama,T.,and Handa,S.: "Characterization of Blood-Group-ABO(H)-Active Glycosphingolipids in Type-AB Human Erythrocytes" Eur.J.Biochem.231. 862-867 (1995)
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[Publications] Ariga,T.,Blaine,M.,G.,Yoshino,H.,Dawson,G.,Kanda,T.,Zeng,G.,Kasama,T.,Kushi,Y.,and Yu,R.Robert: "Glycosphingolipid Composition of Murine Neuroblastoma Cells:O-Acetylesterase Gene Down-Regulates The Expression of O-Acetylated GD3." Biochemistry. 34. 11500-11507 (1995)
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[Publications] Ariga,T.,Bhat,S.,Kanda,T.,Yamawaki,M.,Tai,T.,Kushi,Y.,Kasama,T.Handa,S.,and Yu,K.R.: "Exppression and Localization of Lewis^X Glycolipids and GDla Ganglioside in Human Glioma Cells" Glycoconj.J.12. (1995)
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