1991 年度実績報告書

細胞表層の精密構造認識と機能制御

研究課題

研究課題/領域番号	03236106
研究機関	京都大学
研究代表者	砂本順三京都大学, 工学部, 教授 (80037811)
研究分担者	久枝良雄九州大学, 工学部, 助教授 (70150498) 横山茂之東京大学, 理学部, 教授 (00159229) 猪飼篤東京工業大学, 理学部, 教授 (50011713) 竹市雅俊京都大学, 理学部, 教授 (00025454)
キーワード	血液型抗原 / カドヘレン / αー2ーマクログロブリン / ヒト上皮成長因子 / エンテロバクチンモデル / リポソ-ム / αNーカテニン / B16メラノ-マ
研究概要	本研究班においては、生細胞表面に存在している各種の認識素子、例えば糖脂質または糖蛋白の糖鎖、レセプタ-蛋白のシグナル結合部位、各種抗原性分子とそれに対応する抗体などの認識メカニズムを可能な限り精密構造レベルで追及し、得られた情報・知見を基に実細胞間の情報伝達メカニズムを理解する。本研究では単に化合物間の特異的結合にとどまらず、この精密構造認識機能を通して更に細胞の動的機能の制御へ連継させる点に最大の特長がある。主な成果は次のとうりである。人工境界脂質を含んだリポソ-ムと動物細胞を混合することで、生体膜成分をリポソ-ム膜に構造活性な状態で抽出する方法を確立した。例えば赤血球の血液型抗原やB16メラノ-マの接着蛋白質の移行が観察された(砂本)。Nカドヘリンに結合しているαNーカテニンを同定し、この分子が脳を中心とした神経系に分布していることが発見された。さらに、カテニンのチロシンリン酸化によるカドヘリンの活性抑制が明らかにされた(竹市)。ヒト上皮成長因子(hEGF)とEGFレセプタ-との結合に寄与するアミノ酸残基や高次構造の影響が明らかにされた。さらに蛍光性の非天然型アミノ酸を用いることでhEGFとレセプタ-との動的な認識機構についての知見も得られた(横山)。α2ーマクログロブリンはプロテア-ゼとの結合は非常に小さい活性化エネルギ-と大きな活性化エントロピ-を持つ反応であり、蛋白質内を情報が伝わる速度が以外に遅いことが明らかになった(猪飼)。微生物に含まれるエンテロバクチンのモデル化合物としてマクロ環化合物を合成した。この人工レセプタ-はエンテロバクチンと同様に鉄(III)イオンに対して極めて大きな結合定数を有することが明らかになった。また鉄錯体の絶対配置はカテコ-ル近傍に導入したアミノ酸の立体構造に依存していた(久枝)。

研究成果

(23件)

すべてその他

すべて文献書誌 (23件)

[文献書誌] R.Shibata: "Induction of in vivo and in vitro anti-tulnor responses by sensitization of mice with liposomes containing a crude butanol extract of Leukemia cells and transferred intermembreneously with cell surface protein" Int.J.Cancer. 48. 434-442 (1991)
[文献書誌] Y.Noguchi: "Priming for in vitro and in vivo anti-human T lymphotropic virus type 1 cellular immunity by virus related protein veconstituted into liposome" J.Immunol. 146. 3599-3603 (1991)
[文献書誌] C.J.O'Connor: "Damage of egg phosphatidylcholine liposomes by DNA-binding cytotoxic agents" Bull.Chem.Soc.Jpn.64. 1364-1369 (1991)
[文献書誌] J.Sunamoto: "Direct transfer of tumor surface antigenic Protein from tumor cell to liposome for making liposomal vaccine" J.controlled Release. (1992)
[文献書誌] M.Takeichi: "Cadherin subclasses:Differential expression and their roles in neural morphogenesis" Cold Spring Harbor Quant.Biol.60. 319-325 (1991)
[文献書誌] M.Takeichi: "Cadherin cell adhesion receptors as a morphogentic regulator" Science. 251. 1451-1455 (1991)
[文献書誌] H.Inuzuka: "R-cadherin:A novel Ca^<2+>-dependent cell-cell adhesion molecular expressed in the retina" Neuron. 7. 69-79 (1991)
[文献書誌] A.Nagafuchi: "The 102 kd cadherin-associated protein:Similarity to vinculin and posttranscriptional regulation of expression" Cell. 65. 849-857 (1991)
[文献書誌] H.Inuzuka: "Differential expression of R- and N- cadherin in the nervous system and connective fissues during early chicken development" Development. 113. 959-968 (1991)
[文献書誌] M.Hatta: "Genomic organization and chromosomal mapping of the mouse P-cadherin gene" Nucleic Acis research. 16. 4437-4441 (1991)
[文献書誌] H.Shiozaki: "Expression of immuno-reactive E-cadherin adhesion molecules in human cancers" American J.Pathol. 139. 17-23 (1991)
[文献書誌] S.Hirano: "Dorosophila PS integrin recognize vertebrate vitronedin and function as cell-substrate adhesion receptors in vitro" Development. 113. 1007-1016 (1991)
[文献書誌] S.Ohsako: "Threonin-286 as Autophosphorylation site for appearance of Ca^<2+>-independent activity of Calmodulin-dependent Protein Kinase II α subunit" J.Biochem. 109. 137-143 (1991)
[文献書誌] P.B.Armstrong: "Structure of α_2-macroglobulin from the Arthropod Limulus PolyPhemus" J.Biol.Chem.266. 2526-2530 (1991)
[文献書誌] T.Takahashi: "Electron microscopic and biochemical evidence that proline β-naphtyl-amidase is composed of three identical suburits" FEBS Lett. 280. 297-300 (1991)
[文献書誌] B.Osterberg: "X-ray scattering Study of Hagfish protease inhibifor,a protein structually related to comploment and alpha -2- macroglobulin" Biochemistry. 30. 7873-7878 (1991)
[文献書誌] A.Ikai: "Electron microscopy of 26S complex containing 20S proteasome" FEBS Lett. 292. 21-24 (1991)
[文献書誌] H.Arakawa: "Kinetics of the conformational change of α_2-macroglobulin observed with X-ray solution scattering and fluorescence" Biol.Chem.Hoppe-Seyler. 372. 129-130 (1991)
[文献書誌] J.P.Quigley: "Reaction of Proteinase with α_2-macroglobulin from the american horseshue crab,Limulus," Biol.Chem. 266. 19426-19431 (1991)
[文献書誌] H.Koide: "A site directed mutagenesis study on the role of isuleusine-23 of haman epidermal growth factor in the receptor binding" Biochim.Biophys.Acta. (1992)
[文献書誌] M.Shirouszu: "Glutamic acid residue at position 31 of ras protein is essential to the signal transduction for neurite outgrowth of PC12 cells and the stimulation of GTP ase activity by GAP^<ras>" Oncogen. (1992)
[文献書誌] J.Fujita,Yoshigaki: "Identification of amino acid residue of ras protein that are essential to the signal transducyion for neatrite outgrowth of PC12 cells and the stimulation of GTP ase activity by GAPRas" FEBS Lett. (1992)
[文献書誌] Y.Hisaeda: "Absolute contigurations of Iron (III) complexes of an entero bactin model,a hexapus cyclophane with catechol segments" Chem.lett.1991. 2139-2142 (1991)

1991 年度 実績報告書

細胞表層の精密構造認識と機能制御

研究代表者

砂本 順三 京都大学, 工学部, 教授 (80037811)

研究成果

[文献書誌] R.Shibata: "Induction of in vivo and in vitro anti-tulnor responses by sensitization of mice with liposomes containing a crude butanol extract of Leukemia cells and transferred intermembreneously with cell surface protein" Int.J.Cancer. 48. 434-442 (1991)

[文献書誌] Y.Noguchi: "Priming for in vitro and in vivo anti-human T lymphotropic virus type 1 cellular immunity by virus related protein veconstituted into liposome" J.Immunol. 146. 3599-3603 (1991)

[文献書誌] C.J.O'Connor: "Damage of egg phosphatidylcholine liposomes by DNA-binding cytotoxic agents" Bull.Chem.Soc.Jpn.64. 1364-1369 (1991)

[文献書誌] J.Sunamoto: "Direct transfer of tumor surface antigenic Protein from tumor cell to liposome for making liposomal vaccine" J.controlled Release. (1992)

[文献書誌] M.Takeichi: "Cadherin subclasses:Differential expression and their roles in neural morphogenesis" Cold Spring Harbor Quant.Biol.60. 319-325 (1991)

[文献書誌] M.Takeichi: "Cadherin cell adhesion receptors as a morphogentic regulator" Science. 251. 1451-1455 (1991)

[文献書誌] H.Inuzuka: "R-cadherin:A novel Ca^<2+>-dependent cell-cell adhesion molecular expressed in the retina" Neuron. 7. 69-79 (1991)

[文献書誌] A.Nagafuchi: "The 102 kd cadherin-associated protein:Similarity to vinculin and posttranscriptional regulation of expression" Cell. 65. 849-857 (1991)

[文献書誌] H.Inuzuka: "Differential expression of R- and N- cadherin in the nervous system and connective fissues during early chicken development" Development. 113. 959-968 (1991)

[文献書誌] M.Hatta: "Genomic organization and chromosomal mapping of the mouse P-cadherin gene" Nucleic Acis research. 16. 4437-4441 (1991)

[文献書誌] H.Shiozaki: "Expression of immuno-reactive E-cadherin adhesion molecules in human cancers" American J.Pathol. 139. 17-23 (1991)

[文献書誌] S.Hirano: "Dorosophila PS integrin recognize vertebrate vitronedin and function as cell-substrate adhesion receptors in vitro" Development. 113. 1007-1016 (1991)

[文献書誌] S.Ohsako: "Threonin-286 as Autophosphorylation site for appearance of Ca^<2+>-independent activity of Calmodulin-dependent Protein Kinase II α subunit" J.Biochem. 109. 137-143 (1991)

[文献書誌] P.B.Armstrong: "Structure of α_2-macroglobulin from the Arthropod Limulus PolyPhemus" J.Biol.Chem.266. 2526-2530 (1991)

[文献書誌] T.Takahashi: "Electron microscopic and biochemical evidence that proline β-naphtyl-amidase is composed of three identical suburits" FEBS Lett. 280. 297-300 (1991)

[文献書誌] B.Osterberg: "X-ray scattering Study of Hagfish protease inhibifor,a protein structually related to comploment and alpha -2- macroglobulin" Biochemistry. 30. 7873-7878 (1991)

[文献書誌] A.Ikai: "Electron microscopy of 26S complex containing 20S proteasome" FEBS Lett. 292. 21-24 (1991)

[文献書誌] H.Arakawa: "Kinetics of the conformational change of α_2-macroglobulin observed with X-ray solution scattering and fluorescence" Biol.Chem.Hoppe-Seyler. 372. 129-130 (1991)

[文献書誌] J.P.Quigley: "Reaction of Proteinase with α_2-macroglobulin from the american horseshue crab,Limulus," Biol.Chem. 266. 19426-19431 (1991)

[文献書誌] H.Koide: "A site directed mutagenesis study on the role of isuleusine-23 of haman epidermal growth factor in the receptor binding" Biochim.Biophys.Acta. (1992)

[文献書誌] M.Shirouszu: "Glutamic acid residue at position 31 of ras protein is essential to the signal transduction for neurite outgrowth of PC12 cells and the stimulation of GTP ase activity by GAP^<ras>" Oncogen. (1992)

[文献書誌] J.Fujita,Yoshigaki: "Identification of amino acid residue of ras protein that are essential to the signal transducyion for neatrite outgrowth of PC12 cells and the stimulation of GTP ase activity by GAPRas" FEBS Lett. (1992)

[文献書誌] Y.Hisaeda: "Absolute contigurations of Iron (III) complexes of an entero bactin model,a hexapus cyclophane with catechol segments" Chem.lett.1991. 2139-2142 (1991)

1991 年度実績報告書

砂本順三京都大学, 工学部, 教授 (80037811)