1991 Fiscal Year Annual Research Report

細胞表層の精密構造認識と機能制御

Research Project

Project/Area Number	03236106
Research Institution	Kyoto University
Principal Investigator	砂本順三京都大学, 工学部, 教授 (80037811)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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)。

Research Products
(23 results)

All Other

All Publications (23 results)

[Publications] 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)
[Publications] 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)
[Publications] C.J.O'Connor: "Damage of egg phosphatidylcholine liposomes by DNA-binding cytotoxic agents" Bull.Chem.Soc.Jpn.64. 1364-1369 (1991)
[Publications] J.Sunamoto: "Direct transfer of tumor surface antigenic Protein from tumor cell to liposome for making liposomal vaccine" J.controlled Release. (1992)
[Publications] M.Takeichi: "Cadherin subclasses:Differential expression and their roles in neural morphogenesis" Cold Spring Harbor Quant.Biol.60. 319-325 (1991)
[Publications] M.Takeichi: "Cadherin cell adhesion receptors as a morphogentic regulator" Science. 251. 1451-1455 (1991)
[Publications] H.Inuzuka: "R-cadherin:A novel Ca^<2+>-dependent cell-cell adhesion molecular expressed in the retina" Neuron. 7. 69-79 (1991)
[Publications] A.Nagafuchi: "The 102 kd cadherin-associated protein:Similarity to vinculin and posttranscriptional regulation of expression" Cell. 65. 849-857 (1991)
[Publications] 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)
[Publications] M.Hatta: "Genomic organization and chromosomal mapping of the mouse P-cadherin gene" Nucleic Acis research. 16. 4437-4441 (1991)
[Publications] H.Shiozaki: "Expression of immuno-reactive E-cadherin adhesion molecules in human cancers" American J.Pathol. 139. 17-23 (1991)
[Publications] S.Hirano: "Dorosophila PS integrin recognize vertebrate vitronedin and function as cell-substrate adhesion receptors in vitro" Development. 113. 1007-1016 (1991)
[Publications] 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)
[Publications] P.B.Armstrong: "Structure of α_2-macroglobulin from the Arthropod Limulus PolyPhemus" J.Biol.Chem.266. 2526-2530 (1991)
[Publications] T.Takahashi: "Electron microscopic and biochemical evidence that proline β-naphtyl-amidase is composed of three identical suburits" FEBS Lett. 280. 297-300 (1991)
[Publications] 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)
[Publications] A.Ikai: "Electron microscopy of 26S complex containing 20S proteasome" FEBS Lett. 292. 21-24 (1991)
[Publications] 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)
[Publications] J.P.Quigley: "Reaction of Proteinase with α_2-macroglobulin from the american horseshue crab,Limulus," Biol.Chem. 266. 19426-19431 (1991)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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 Fiscal Year Annual Research Report

細胞表層の精密構造認識と機能制御

Principal Investigator

砂本 順三 京都大学, 工学部, 教授 (80037811)

Research Products

[Publications] 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)

[Publications] 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)

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

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

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

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

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

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

[Publications] 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)

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

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

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

[Publications] 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)

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

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

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

砂本順三京都大学, 工学部, 教授 (80037811)