糖鎖発現の制御因子

• Project/Area Number: 05274107
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Research Institution: Tokyo Metropolitan Organization for Medical Research
• Principal Investigator: 鈴木 明身 財団法人 東京都臨床医学総合研究所, 生体膜研究部門, 研究員 (70134533)
• Co-Investigator(Kenkyū-buntansha): 菅原 一幸 神戸薬科大学, 薬学部, 教授 (60154449) ; 伊東 信 九州大学, 農学部, 助教授 (40253512) ; 中田 博 京都産業大学, 工学部, 教授 (90113141) ; 羽渕 脩躬 愛知教育大学, 教育学部, 教授 (90024067) ; 岩森 正男 東京大学, 医学部, 助教授 (90110022) ; 飯田 静夫 東京医科歯科大学, 医学部, 教授 (00009987) ; 松本 勲武 お茶の水女子大学, 理学部, 教授 (70012664) ; 西川 義尚 東海大学, 工学部, 教授 (20109953) ; 牧田 章 北海道大学, 癌研生化学, 教授 (60004561)
• Project Period (FY): 1993 – 1996
• Project Status: Completed (Fiscal Year 1996)
• Budget Amount: ¥169,200,000 (Direct Cost: ¥169,200,000); Fiscal Year 1996: ¥41,200,000 (Direct Cost: ¥41,200,000); Fiscal Year 1995: ¥42,000,000 (Direct Cost: ¥42,000,000); Fiscal Year 1994: ¥45,000,000 (Direct Cost: ¥45,000,000); Fiscal Year 1993: ¥41,000,000 (Direct Cost: ¥41,000,000)
• Keywords: N-グリコリルノイラミン酸 / CMP-NeuAc水酸化酵素 / セラミド / エンドグリコセラミダーゼ / 硫酸転移酵素 / シアリルルイスA / プロテオグリカン / ビトロネクチン / CMP-NeuAc水酸化酸素 / ギラン・バレー症候群 / がん関連抗原 / シアリルLe^a抗原 / レクチン / ガラクトース転移酵素 / Tn抗原 / アネキシンファミリー

Research Abstract

N-グリコリルノイラミン酸(NeuGc)の発現を律速制御していると考えられるCMP-NeuAc水酸化酵素の解析を行い,ヒトcDNAをクローニングした.COS細胞に強制発現させても活性は検出されず,ヒトでNeuGcの発現抑制される原因と考えられた.マウスβ-1,6-GlcNAc転移酵素のゲノムの構築が明らかになり,組織特異的発現制御の関与が示された(鈴木).乳腺腺構造分化に伴って,硫酸転移酵素と2-6シアル酸転移酵素の活性化と2-3シアル酸転移酵素の抑制が起こった.硫酸転移酵素はEGF,2-6シアル酸転移酵素はプロラクチンが関与する(岩森).ヒト腎癌細胞から糖脂質硫酸転移酵素を精製し,アミノ酸配列に基づいてcDNAをクローニングすることに成功した.新しい構造の酵素であり,増殖因子で酵素が誘導されることから,機序の解明が可能になったった(本家).コンドロイチン6硫酸転移酵素cDNAをCOS7細胞に発現させるとガラクトースの6位にも硫酸化が起こった(羽渕).グリコスアミノグリカンの蛋白質への結合領域に,糖鎖伸長の仕訳機能を持つ新規構造,および新規α-GalNAc転移酵素を見いだした(菅原).糖鎖とセラミドを切断する酵素を使い,グルコース転移酵素が糖脂質発現の律速となる事を見いだした.セラミドN-アシラーゼを同定し,糖鎖工学へ応用した(伊東).O-アセチルセラミドを持つGM3の合成法を確立し,抗体を作製した(賀佐).ガングリオシドとスルファチドがDNA polymeraseαを抑制する.スフィンゴシンはRNA primaseを阻害することが明らかになった(瀧).アドリアマイシン耐性株で糖脂質に変化が見られた(保田).TPA処理によるヒト大腸癌細胞の糖鎖の変化は,より多くの糖鎖の付加に原因がある事が明らかになった(中田).新型レクチンのアネキシンIVの構造,cDNAの塩基配列を明らかにした.リン脂質が糖結合に影響を与える事が明らかになった.(松本).

Research Products

• [Publications] Y.Hashimoto,A.Oshima,H.Narimatsu,& A.Suzuki: "Expression of the cholera toxin B subunit in the Glogi apparatus of Swiss 3T3 cells inhibits DNA synthesis induced by basic fibroblast growth factor." J.Biochem.119. 985-868 (1996)
• [Publications] K.Honke,M.Tsuda,Y.Hirahara,Hirahara,A.Ishii et al..: "Molecular cloning and expression of cDNA encoding human 3′-phosphoadenylylsulfate:galactosylceramide 3′-sulfotransferase." J.Biol.Chem.272. 4864-4868 (1997)
• [Publications] O.Habuchu,Y.Hirahara,K.Uchimura,& M.Fukuta.: "Enzymatic sulfation of galactose residue of keratan sulfate by chondroitin 6-sulfotransferase." Glycobiology. 6. 51-57 (1996)
• [Publications] H.Kitagawa,M.Ujikawa: "Developmental changes in serum UDP-GlcA:chondroitin glucuronyltransferase activity." J.Biol.Chem.271. 6583-6585 (1996)
• [Publications] M.Ito,& H.Komori: "Homeostasis of cell-surface glycosphingolipid content in B16 melanoma cells.-Evidence revealded by an endoglycoceramidase-" J.Biol.Chem.271. 12655-12660 (1996)
• [Publications] K.Kojima,K.Yamamoto,T.Irimura,T.Osawa,I.Ogawa et al.: "Characterization of carbohydrate-binding protein p33/41. Relation with annexin IV,molecular basis of the doublet forms (p33/410,and modulation of the carbohydrate binding avtivity by phospholipids." J.Biol.Chem. 271. 7679-7885 (1996)
• [Publications] Kawano, T.: "Molecular cloning of cytidine monophospho-N-acetylneuraminicacid hydroxylase." J. Biol. Chem.270. 16458-16463 (1995)
• [Publications] Hirota, K.: "Alteration in the reactivity of sphingomyelin in mitogen-stimulated lymphocytes." J. Biochem.118. 4-8 (1995)
• [Publications] Ito, M.: "A novel enzyme that cleaves the N-acyl linkage of ceramides in various glyco-sphingolipids as well as sphingomyelin to produce their lyso forms." J. Biol. Chem.270. 24370-24374 (1995)
• [Publications] Ji, L.: "The hydrolysis of cell surface glycosphingolipids by endoglycoceramidase reduces epidermal growth factor receptor phosphrylation in A431 cells" Glycobiology. 5. 343-350 (1995)
• [Publications] Nakada, H.: "Expression of the antigen on a T-lymphoid cell line, SupT1." Glycconjugates J.12. 356-359 (1995)
• [Publications] Kitagawa, H.: "GalNAc transfer to the common carbohydrate-protein linkage region of sulfated glycos-aminoglycans." J. Biol. Chem.270. 22190-22195 (1995)
• [Publications] T.KAWANO: "Biosynthesis of N-glycolyneuraminic acid-containing glycoconjugates.Purification and characterization of the key enzyme of the cytidine monopfospho-N-acetyl-heuraminic acid hydroxylation system." J.Biol.Chem.269. 9024-9029 (1994)
• [Publications] H.Takematsu: "Reaction mechanism underlying CMP-N-acetylneuraminicacid hydroxyiation in mouse liver:Formation of a ternary complex of cyto-chrome b5,CMP-N-acetylneuramiuic acid,and a hydroxylation enzyme." J.Biochem.115. 381-386 (1994)
• [Publications] M.Sekine: "Purification and characterization of UDP-GlcNAC:IV^3βGal-Gb_4Cer β-1,6-GIcNAc transferase from mouse kidney." J.Biol.Chem.269. 31143-31148 (1994)
• [Publications] K.Nakamura: "IV^3α(NeuGcα2-8NeuGc)-g_<g4> cer is restricted to cD4^γT cells producing interleukin-2 and a small population of mature thymocytes in mice" J.Biol.Chem.270. 3876-3881 (1995)
• [Publications] N.Yuki: "molecular mimicry between GQ16 ganglioside and lipopoly saccharides of campylovacter jejuni isolated from patients with Fishers syndrome." Ann.Neurol.36. 791-793 (1994)
• [Publications] M.Kagehara: "Programmed expression of cholesterol sulfotransferase and transglutaminase during epidermal differentiation of marine skin olevelopment." Biochim.Biophys.Acta. 1215. 183-189 (1994)
• [Publications] Kawano,T.et al.: "Regulation of biosynthesis of N-glycolylneuraminic acid-containing glycoconjugates:characterization of factors required for NADH-dependent CMP-NeuAc hydroxyation." Glycoconjugate J.10. 109-115 (1993)
• [Publications] Hashimoto,Y.et al.: "Purification and characterization of UDP-N-acetylgalactosamine:GM3/GD3 N-acetylgalactosaminyltransferase(EC2.4.1.92)from mouse liver." J.Biol.Chem.268. 25857-25864 (1993)
• [Publications] Kawano,T.et al.: "Biosynthesis of N-glycolylneuraminic acid-containing glycoconjugates:Purification and characterization of the key enzyme of the cytidine monophospho-N-acetylneuraminic acid hydroxylation system." J.Biol.Chem.269. 9024-9029 (1994)
• [Publications] Kobayashi,T.etal: "Sulfolipids and glycolipid sulfotransferase activities in human renal cell carcinoma cells." Brit.J.Cancer. 67. 76-80 (1993)
• [Publications] Kobayashi,T.et al.: "Involvement of protein kinase C in the regulation of glycolipid sulfotransferase activity levels in renal cell carcinoma cells" Cancer Res.53. 2484-2489 (1993)
• [Publications] Kobayashi,T.et al.: "Epidermal growth factor elevates the activity of glycolipid sulfotransferase in renal cell carcinoma cells." Int.J.Cancer.55. 448-452 (1993)
• [Publications] Kobayashi,T.et al.: "Cell surface sulfoglycolipids are involved in the attachment of renal cancer cells to laminin." Int.J.Cancer. 56. 281-285 (1994)
• [Publications] Yoshino,H.et al.: "Fucosyl-GM1 in human sensory nervous tissue is a target antigen in patients with autoimmune neuropathies." J.Neurochem.61. 658-663 (1993)
• [Publications] Matsuda,K.et al.: "Glycosphingolipid compositions of human T-lymphotropic virus type I (HTLV-I )and human immunodeficiency virus(HIV)-infected cell lines." Biochem.Biophys.Acta. 1168. 123-129 (1993)
• [Publications] Yuki,N.et al.: "A bacterium lipopolysaccharide that elicits Guillan-Barre syndrome has a GM1 gaglioside like structure." J.Exp.Med.178. 1771-1775 (1993)
• [Publications] Cui,Y.et al.: "Human cervical epidermal carcinoma-associated intracellular localization of glycosphimgolipid with blood group type 3 chain." Jpn.J.Cancer Res.84. 664-672 (1993)
• [Publications] Iwamori,M.et al.: "Compensatory occurrence of IV2Fuc,II3NeuAc-Gg4Cer and human fetal antigen,Lc4Cer,in small cell carcinoma of human lung." Cancer Res.53. 5903-5907 (1993)
• [Publications] Kitagawa,H.et al.: "Novel oligosaccharides with the sialy-Lea structure in human milk." J.Biochem.114. 504-508 (1993)
• [Publications] Nakada,H.et al.: "Epitopic structure of Tn glycophorin A for an anti-Tn antibody(MLS128)." Proc.Natl.Acad.Sci.USA. 90. 2495-2499 (1993)
• [Publications] Kitagawa,H.et al.: "Novel gangliosides containing the sialyl-Lea structure from a human rectal adenocarcinoma." J.Biol.Chem.268. 26541-26545 (1993)