ガングリオシド糖鎖認識プロ-ブの開発

1991 Fiscal Year Annual Research Report

• Project/Area Number: 03255103
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 飯田 静夫 東京医科歯科大学, 医学部, 教授 (00009987)
• Co-Investigator(Kenkyū-buntansha): 谷口 直之 大阪大学, 医学部, 教授 (90002188) ; 山形 達也 三菱化成生命科学研究所, 細胞認識研究部, 部長 ; 山科 郁男 京都産業大学, 工学部, 教授 (70025675) ; 神奈木 玲児 愛知県がんセンター研究所, 部長 (80161389) ; 内貴 正治 国立予防衛生研究所, 獣疫部, 部長 (10020752)
• Keywords: 糖脂質 / ガングリオシド / モノクロ-ナル抗体 / 糖転移酵素 / エンドグリコシダ-ゼ / がん / 細胞接着因子 / 糖鎖

Research Abstract

ガングリオシド認識のプロ-ブとして結合タンパク質、抗体、糖の分解・転移酵素の研究を行って以下の成果を得た。
1。豚新生仔下痢症を起こす病原性大腸菌の繊毛はセラミド部分に水酸基をもつGM3(NeuGc)の結合親和性が高く、この糖脂質は成長に伴い減少することを見いだした(内貴)。2。シアロ糖鎖に特異的なモノクロ-ナル抗体を作成し、リンパ球系のシアリルSSEAー1はNK細胞と抗原刺激を受け活性化されたT細胞にのみ発現しており、消化器癌ではシアリルLe^aが強く発現し、両者ともELAMー1に結合する細胞性接着因子であることを明らかにした(神奈木)。結腸癌細胞と反応する抗体MSW113の抗原決定基糖鎖はシアリルLe^a構造であるが、NSー19ー9とは異なる抗原認識部位を持つ。この抗体を用いてヒト結腸癌組織から新しい4種の糖脂質を同定した。またこの組織のムチンには抗原決定基を持つ分子サイズの異なる多種類の糖鎖が存在していた(山科)。3。糖脂質の糖鎖部分を切り放すエンドグリコシダ-ゼを精製アクチベ-タ-と共に細胞に作用させると可逆的に増殖が抑制され、EGFのレセプタ-との結合は変化しないが、レセプタ-のリン酸化は低下することを見いだした(山形)。肝臓でグルコサミン転移酵素III活性もγGTPも非実質細胞群に高く、また糖鎖の分析から非実質細胞群由来のγGTPにはbisecting GlcNAcが存在することを明らかにした(谷口)。ガラクト-ス転移酵素活性は癌患者血清中に高値であるが、培養癌細胞から培地中にやや分子量の小さい酵素が放出されていることを見いだした(飯田)。4。前口動物の中で、軟体動物の二枚貝にマンノ脂質、巻貝類にガラクト脂質、また節足動物にはマンノ脂質が主成分で、さらに環形動物にガラクト脂質と新型ホスホコリン含有糖脂質の存在を明らかにした(杉田)。

Research Products

• [Publications] Takao Taki,et al.: "Had-1,a Uridine 5'-Diphosphogalactose Transport-defective Mutant of Mouse Mammary Tumor Cell FM3A:Composition of Glycolipids,Cell Growth Inhibition by Lactosylceramide,and Loss of Tumorigenicity" Cancer Res.51. 1701-1707 (1991)
• [Publications] Koh Asano,et al.: "Acidic Glycosphingolipids of Human Amnion" J.Biochem.109. 106-112 (1991)
• [Publications] Takeshi Kasama,et al.: "Structural Studies of Gangliosides by Fast Atom Bobmbardment Ionization,Low-Energy Collision-Activated Dissociation,and Tandem Mass Spectrometry" Biochemstry. 30. 5621-5624 (1991)
• [Publications] M.Midorikawa,et al.: "Uptake and Metabolism of Radiolabelled GMl-ganglioside in Skin Fibroblasts from Controls and Patients with GMl-gangliosidosis" J.Inher.Metab.Dis.14. 721-729 (1991)
• [Publications] Shigeru Nishiwaki,et al.: "Elevations of β l-4galactosyltransferase activity in sera of patients with cancer" Cacer Res.(1992)
• [Publications] Takao Taki,et al.: "13C-Nuclear magnetic resonance of glycosphingolipids" J.Biochem.(1992)
• [Publications] Takao Taki,et al.: "Human meconium gangliosides - Characterization of a novel I-type ganglioside with NeuAc α 2-6Gal structure" J.Biol.Chem.(1992)
• [Publications] Yuyama,Y.: "Postnatal change of pig intestinal ganglioside bound by Escheriehia coli with K99 fimbriae" J.Biol.Chem.
• [Publications] Mori,Y.: "Genetic studies on experimental autoimmune gastritis induced by neonatal thymectomy using recombinant inbred strains between a high-incidence strain,BALB/c,and a low-incidence strain,DBA/2." Clin,Exp.Immunol.84(1). 145-152 (1991)
• [Publications] Ando,Y.: "Calcium-induced intracellular cross-linking of lipocortin I by tissue transglutaminase in A431 cells: Augmentation by membrane phospholipids." J.Biol.Chem.266. 1101-1108 (1991)
• [Publications] 神奈木 玲児,銭田 晃一,恒松 徳五郎: "「自己免疫性溶血性貧血における糖鎖性の自己抗原」" 臨床血液. 32(6). 599-605 (1991)
• [Publications] 神奈木 玲児: "「血液型物質・糖鎖抗原・腫瘍マ-カ-」" 生物物理化学. 35(6). 421-426 (1991)
• [Publications] Hiroshi Nakada: "Expression of the Tn antigen on T-lymphoid cell line Jurkat" Biochem.Biophys.Res.Commun.179-2. 762-767 (1991)
• [Publications] Kazuyuki Sugahara: "Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chon-droitin sulfate proteoglycans of whale cartilage" Eur.J.Biochem.202-3. 805-811 (1991)
• [Publications] 小野 恵利子: "毒素原性大腸菌K99線毛アドヘシンの検索" 家蓄生化学研究会報. 26. 1-11 (1991)
• [Publications] 内貴 正治: "細菌やウイルスが見分ける細胞の顔" 現代化学. 11. 38-42 (1991)
• [Publications] One,E.: "The nucleotide sequense of the genes,fanE and fanF of Escherichia coli K99 fimbriae" Jpn.J.Vet.Res.39. 1-10 (1991)
• [Publications] Yoshimatsu,K.: "New methods for isolation of K99 fimbriae from enterotoxigenic Escherichia coli(NOTE)" J.Vet.Med.Sci.53. 1119-1121 (1991)
• [Publications] Nakao,H.: "Hypogalactosylation of immunoglobulin G sugar chains and elevated serum interleukin 6 in Castleman's disease" Clin.Chim.Acta. 197. 221-228 (1991)
• [Publications] Kinoshita,N.: "Glycosylation at the Fab portion of myeloma immunoglobulin G and increased fucosylated biantennary sugar chains:Structural analysis by HPLC and antibody-lectin enzyme immunoassay using lens culinaris agglutinin." Cancer Research. 51. 5888-5892 (1991)
• [Publications] Gu,J.: "Biosynthesis of blood group I and i antigens in rat tissues: Identification of a novel β1-6-N-acetylglucosaminyltransferase" J.Biol.Chem.267. 2994-2999 (1992)
• [Publications] Ohno,M.: "Enzymatic basis of sugar structures of α-fetoprotein in hepatoma and hepatoblastoma cell lines: Correlation with activities of α1-6 fucosyltransferase and N-acetylglucosaminyltransferases III and V." Int.J.Cancer. (1992)
• [Publications] Takada,A.: "Adhesion of human cancer cells to vascular endotherium mediated by a carbohydrate antigen,sialyl Lewis A" Biochem.Biophys.Res.Commun.179. 713-719 (1991)
• [Publications] Itai,S.: "Tissue distribution of sialyl 2-3 and 2-6 Lewis A antigen ratio for the digestive tract" Cancer. 67. 1576-1587 (1991)
• [Publications] Shigeyuki Fukui: "Characterization of mucin-antigens recognized by monoclonal antibodies raised against human colon cancer cells" Cancer Research. 51-1. 331-335 (1991)
• [Publications] Hiroshi Kitagawa: "Novel oligosaccharides with the sialyl-Le^a structure in human milk" Biochemistry. 31-11. 2869-2876 (1991)
• [Publications] Masahiro Maeda: "Expression of cDNA for batroxobin,a thrombin-like snake venom enzyme" J.Biochem.109-4. 632-637 (1991)
• [Publications] Hiroshi Nakada: "Elucidation of an essential structure recognized by an anti-Tn monoclonal antibody (MLS 128)" J.Biol.Chem.266-19. 12402-12405 (1991)
• [Publications] Hiroshi Kitagawa: "Characterization of mucin-type oligosaccharides with the sialyl-Le^a structure from human colorectal adenocarcinoma cells" Biochem.Biophys.Res.Commun.178-3. 1429-1436 (1991)
• [Publications] Hiroshi Kitagawa: "Isolation and structural studies of human milk oligosaccharides that are reactive with a monoclonal antibody MSW 113" J.Biochem.110-4. 598-604 (1991)
• [Publications] 杉田 陸海: "ウロン酸糖脂質の免疫化学的研究ー抗IgM Mー蛋白質結合性グルクロン酸糖脂質抗体の調整ー" 滋賀大学教育学部紀要. 41. 33-40 (1992)
• [Publications] 板坂 修: "頭足類の糖脂質(I)スルメイカの中性糖脂質" 滋賀大学教育学部紀要. 41. 41-46 (1992)
• [Publications] 杉田 陸海: "環形動物の複合糖脂質ーヒトツモンミミズPheretina hilgendorfiの中性糖脂質ー" 油化学. 41. (1992)
• [Publications] 杉田 陸海: "節足動物の新しい糖脂質" 蛋白質核酸酵素. (1992)
• [Publications] Itonori,S.: "Characterization of a new phosphonocerebroside,N-methyl-2-aminoethyl-phosphonylglucosylceramide.from the antarctic krill,Euphausia superba." Biochim.Biophys.Acta. 1081. 321-327 (1991)
• [Publications] 杉田 陸海: "ホスホノ脂質の機能ー糖脂質抗体と糖脂質の反応におけるホスホノ脂質(2ーアミノエチルホスホン酸セラミド)の効果" 滋賀大学教育学部紀要. 40. 35-39 (1991)
• [Publications] 杉田 陸海: "マンノ脂質の免疫化学的研究(酵素抗体法によるaーマンノシダ-ゼ活性の測定)" 油化学. 40. 427-431 (1991)
• [Publications] 堀 太郎: "マンノ脂質の免疫化学的研究ー抗ceramide nonasaccharide抗体の性状および貝類でのハプテンの検出ー" 滋賀文化短期大学紀要. 1. 1-7 (1991)
• [Publications] Itonori,S: "Polar glycosphingolipids in insect: Chemical structures of glycosphingo-lipid series containing 2' -aminoethylphosphoryl(-6)-N-acetylglucosamine as a polar group from larvae of the green-bottle fly,Lucilia caesar" J.Biochem.110. 479-485 (1991)
• [Publications] Itonori,S: "Immunogenic properties of mannose-containing ceramide disaccharide and immunochemical detection of its hapten in the two kinds of crustacean.Euphausia superba and Macrobrachium nipponense." Biochimica Biophysica Acta. 1089. (1992)
• [Publications] Ito,M.,et al.: "Activator proteins for glycosphingolipid hydrolysis by endoglycoceramidase: Elucidation of biological functions of cell-surface glycosphingolipids in situ by endoglycoceramidases made possible using these activator proteins." J.Biol.Chem.266. 7919-7926 (1991)
• [Publications] Ito,M.,et al.: "Conversion of endoglycoceramidase-activator II by trypsin to the 27.9kD polypeptide possessing full activity:Purification of activator for endoglycoceramidase by trypsin treatment followed by trypsin inhibitor agarose column application." J.Biochem.110. 328-332 (1991)
• [Publications] Shimamura,M.,et al.: "The retarded rate of acid-catalyzed solvolysis of glycoside bonds between reducingend glucose residue and ceramide in glycosphingolipids compared with that of glycoside bonds between hexopyranosides." FEBS Letters. 290. 213-215 (1991)
• [Publications] Yamagata,T.and Yamazaki,K.: "Implanting mouse embryos stain with a LNF-I bearing fluorescent probe at their mural trophectodermal side." Biochem.Biophys.Res.Commun.181. 1004-1009 (1991)
• [Publications] Higashi,H.,and Yamagata,T.: "Calmodulin,A ganglioside-binding protein." J.Biol.Chem.
• [Publications] Higashi,H.,et al.: "Mechamism for ganglioside-mediated modulation of calmodulin-dependent enzyme." J.Biol.Chem.
• [Publications] Shimamura,M.,et al.: "Presence of murine fetal liver cells capable of being induced to differentiate in vitro into T cell receptor α β positive cells." Eur.J.Immunol.
• [Publications] Shimamura,M.,et al.: "Hematopoietic cell lines capable of colonizing the thymus following in vivo transfer expressed T cell receptor δ gene immature mRNA." J.Exp.Med.
• [Publications] 山形 達也: "細胞表層の糖鎖生物学ー糖鎖工学の芽生えー" バイオサイエンスとインダストリ-. 49. 1055-1065 (1991)
• [Publications] 山形 達也: "T細胞初期分化研究のための細胞生物学的アプロ-チ" ガングリオシドニュ-ス. 2(3). 7-15 (1991)
• [Publications] 伊東 信: "エンドグリドセラミダ-ゼを使った細胞表層糖脂質の機能解析" 蛋白質・核酸・酵素 特集号. (1992)
• [Publications] 山形 達也: "着床における糖鎖の役割" 蛋白質・核酸・酵素 特集号. (1992)
• [Publications] 神奈木 玲児: "「臓器移植と赤血球の糖鎖抗原」 "ABO血液型不適合間賢移植"" 太田 和夫監修,日本医学館,東京, 7 (1991)
• [Publications] 神奈木 玲児: "「モノクロ-ナル抗体による腫瘍抗原の測定」 "注目の臨床実験検査法"" 中山書店,東京, 15 (1991)
• [Publications] 杉田 陸海(分担執筆): "生物薬科学実験講座 I巻生体構成成分と高分子" 廣川書店,
• [Publications] 山形 連也: "糖鎖工学" 産業調査会, (1992)
• [Publications] YAMAGATA,T.: "Handbook of Endoglycosidases and Amidases" CRC Press,Inc.Florida, (1992)