1995 年度実績報告書

イオン駆動型トランスポータとチャネルの構造・機能協関

研究課題

研究課題/領域番号	07276102
研究機関	岡山大学
研究代表者	土屋友房岡山大学, 薬学部, 教授 (80012673)
研究分担者	野間昭典京都大学, 医学部, 教授 (00132738) 清野進千葉大学, 医学部, 教授 (80236067) 北里宏滋賀医科大学, 医学部, 教授 (20079700) 笠原道弘帝京大学, 医学部, 教授 (40010102) 乾賢一京都大学, 医学部, 教授 (70034030)
キーワード	イオン / トランスポータ / チャネル / 構造と機能
研究概要	代表者・分担者それぞれが開拓し解析しているイオン駆動型トランスポータとイオンチャネルにつき、構造面と機能面から分子レベルでの解析(分子構造、分子機序)を行った。代表者は主として大腸菌のメリビオース輸送系(Na^+/基質シンポータ)とNa^+/H^+アンチポ-タについて遺伝子操作技術による解析により、イオン認識部位・基質認識部位の同定を行った。また、新たにいくつかのNa^+共役型トランスポータの遺伝子クローニングに成功した。一方、笠原と協力し、一次構造上良く似た輸送系同士のキメラを細胞内相同組換えにより作成し、基質認識部位の同定を行った。また、笠原はラット消化管におけるNa^+駆動型グリコース輸送体の局在を明らかにすると共に、酵母細胞での発現系を構築した。乾は、PCR法及び卵母細胞発現系を用いて薬物トランスポータ群及び輸送調節因子のcDNAクローニングと機能解析を行った。野間は心筋のNa^+/Ca^<2+>交換輸送タンパク質について、それぞれのイオンの輸送の速度論的解析を行うと共に、遺伝子面からの解析により、イオン認識部位、調節部位の同定を行った。特に、塩基性アミノ酸残基の多いループ領域の役割を明らかにした。北里は、膵β細胞においてグルコースによって惹起される律動的なスパイクバーストに対するNa^+、Ca^<2+>の影響を解析した。清野は膵β細胞で発現されるイオンチャネルについて解析し、新たなK^+チャネルファミリーメンバーのBIRをクローニングすると共にスルホニル尿素受容体SURとを細胞内で再構成した。そしてATPによりこのK^+チャネルが制御されることを見いだした。この発見はグルコースがインスリン分泌を引き起こすメカニズムの分子基盤を知る上で極めて重要な発見である。

研究成果

(34件)

すべてその他

すべて文献書誌 (34件)

[文献書誌] Inoue,H.: "Essential aspartic acid residues,Asp-133,Asp-163 and Asp-164,in the transmembrane helices of a Na^+/H^+ antiporter (NhaA) from E.coll." FEBS Letters. 363. 264-268 (1995)
[文献書誌] Ohtani,K.: "Involvement of a protein kinase C in Ca2^+-signaling pathways to activation of AP-1 DNA-binding activity evoked via NMDA-" J.Neurochem.65. 605-614 (1995)
[文献書誌] Tomita,Y.: "Transport of oral cephalosporins by the H^+/dipeptide cotransporter and distribution of the transport activity in isolated rabbit intesinal" J.Pharmacol.Exp.Ther.272(1). 63-69 (1995)
[文献書誌] Matsumoto,S.: "Transport characteristics of ceftibuten,a new cephalosporin antibiotic,via the apical H^+/dipeptide cotransport system in the human intestinal" Pharm.Res.12. 1483-1487 (1995)
[文献書誌] Saito,H.,: "Cloning and characterization of a rat H^+/peptide cotransporter mediating absorptoption of β-lactam antibiotics in the intestine" J.Pharmacol.Exp.ther.275(3). 1631-1637 (1995)
[文献書誌] Nagai,J.: "Specificity of p-aminohippurate transport system in the OK kidney epithelial cell line" J.Pharmacol.Exp.ther.274(3). 1161-1166 (1995)
[文献書誌] Hirai,M.: "Cepharantin,a multidrug resistant modifier,is a substrate for P-glycoprotein." J.Pharmacol.Exp.Ther.275(1). 73-78 (1995)
[文献書誌] Yoshida,A.: "Immunohistochemical localization of Na^+-dependent glucose trasnporter in the rat digestive tract." Histochmical J.27. 420-426 (1995)
[文献書誌] Nishizawa,K.: "Substrate recognition domain of the Gal2 galactose transporter in yeast Saccharomyses cerevisiae as revealed by chimeric galactose-" J.Biol.Chem.270. 2423-2426 (1995)
[文献書誌] Takigawa,T.: "Antibodies against GM1 ganglioside affect K^+ and Na^+ currents in isolated rat myelinated nerve fibers." Ann Neurology. 37. 436-442 (1995)
[文献書誌] Guo,J.: "A sustained inward current activated at the diastolic potential range in rabbit sino-atrialnode cells." J.Physiology. 483. 1-13 (1995)
[文献書誌] Manabe,K.: "Hyperpolarization induced by vasoactive substances in intact guinea-pig endocardial endothelial cells." J.Physiology. 484. 25-40 (1995)
[文献書誌] Manabe,K.: "Classification of ion channels in the luminal and abluminal membranes of guinea-pig endocardial endothelial cells." J.Physiology. 484. 41-52 (1995)
[文献書誌] Manabe,K.: "Non-selective cation current of guinea-pig endocardial endothelial cells." J.Physiology. 487. 407-419 (1995)
[文献書誌] Inagaki,N.: "Reconstitution of lKATP : An inward rectifier subunit plus the sulfonylurea receptor." Science. 270. 1166-1170 (1995)
[文献書誌] Inagaki,N.: "cDNA sequence,gene structure,and chromosomal localization of the human ATP-sensitive potassium channel,uKATP-1,Gene (KCNJ8)." Genomics. 30. 102-104 (1995)
[文献書誌] Yamada,Y.: "Human gastric inhibitory polyeptide receptor : cloning of gene and cDNA." Genomics. 29. 773-776 (1995)
[文献書誌] Kuromi,H.: "A monitor of secretion from single pancreatic β-cells." Diabetes. 44. 1213-1217 (1995)
[文献書誌] Yamada,Y.: "The structures of the human calcium channel α1 subunit (CACNL1A2) and β subnuit (CACNLB3) genes." Genomics. 27. 312-319 (1995)
[文献書誌] Marqueze,B.: "Cellular localization of Synaptotagmin I,II,and III mRNAs in the central nervous system,pituitary and adrenal glands of the rat." J.Neurosci.15. 4906-4917 (1995)
[文献書誌] Inagaki,N.: "Expression and role of ionotropic glutamate recepters in pancreatic islet cells." FASEB.J. 9. 686-691 (1995)
[文献書誌] Inagaki,N.: "Cloning and functional characterization of a novel ATP-sensitive potassium channel ubiquitously expressed in rat tissues,including" J.Biol.Chem.270. 5691-5695 (1995)
[文献書誌] Blondel,O.: "Inositol 1,4,5-trisphosphate recetpors,secretory grnules and secretion in endocrine and neuroendocrine cells." Trends Neurosci. 18. 157-161 (1995)
[文献書誌] Yorimitsu,K.: "Cloning and sequence of a human endothelin converting enzyme in renal adenocarcinoma (ACHN) cells producing endothelin-2." Biochem.Biophys.Res.Commun.208. 721-727 (1995)
[文献書誌] Jones,J.M.: "Synaptotagmin genes on mouse Chromosomes 1,7 and 10 and human Chromosome 19." Nammal.Genome. 6. 212-213 (1995)
[文献書誌] Oho,C.: "Expression and complex formation of soluble N-ethyl-maleimide-sensitive factor attachment protein (SNAP) receptors in clonal rat" Neurosci.Lett.186. 208-210 (1995)
[文献書誌] Seino,S: "CACN4,the major α1 subunit isoform of voltage-dependent calsium channels in pancreatic β-cells : α minireview of current progress." Diabetes Res.Clin.Pract.28. S99-S103 (1995)
[文献書誌] lhara,Y.: "Molecular diversity and functional characterization of voltage-dependent calcium channels (CACN4) expressed in pancreatic β-cells." Mol.Endocrinol.9. 121-130 (1995)
[文献書誌] Shin,Y.: "Reconstitution of the F_1-ATPase activty from purified α、β、γ and δ or ε subunits with glutathions S-transferase fused at their amino" Biochim.Biophys.Acta. 1273. 62-70 (1996)
[文献書誌] Sarker,R.l.: "Properties of a Na+/galactose (glucose) symport system in Vibrio parahaemolyticus." Biochim.Biophys.Acta. (in press).
[文献書誌] Sarker,R.l: "Sequence of a Na+/glucose symporter gene and its flanking regions of Vibrio parahaemolyticus." Biochim.Biophys.Acta. (in press).
[文献書誌] Kasahara,T.: "Expression of the rat GLUT1 glucose transporter in the yeast Saccharomyces cerevisiae." Biochemical.J.(in press).
[文献書誌] Ding W-G.: "A possible role of ATP-sensitive K^+ channel in determining the duration of spike-bursts in mouse pancreatic β-cells." Biochim.Biophys.Acta. (in press).
[文献書誌] He L-P.: "Glucagon induced Ca^<2+>-dependent increase of reduced pyridine nucleotides in mouse pancreatic β-cells." Biochim.Biophys.Acta. (in press).

1995 年度 実績報告書

イオン駆動型トランスポータとチャネルの構造・機能協関

研究代表者

土屋 友房 岡山大学, 薬学部, 教授 (80012673)

研究成果

[文献書誌] Inoue,H.: "Essential aspartic acid residues,Asp-133,Asp-163 and Asp-164,in the transmembrane helices of a Na^+/H^+ antiporter (NhaA) from E.coll." FEBS Letters. 363. 264-268 (1995)

[文献書誌] Ohtani,K.: "Involvement of a protein kinase C in Ca2^+-signaling pathways to activation of AP-1 DNA-binding activity evoked via NMDA-" J.Neurochem.65. 605-614 (1995)

[文献書誌] Tomita,Y.: "Transport of oral cephalosporins by the H^+/dipeptide cotransporter and distribution of the transport activity in isolated rabbit intesinal" J.Pharmacol.Exp.Ther.272(1). 63-69 (1995)

[文献書誌] Matsumoto,S.: "Transport characteristics of ceftibuten,a new cephalosporin antibiotic,via the apical H^+/dipeptide cotransport system in the human intestinal" Pharm.Res.12. 1483-1487 (1995)

[文献書誌] Saito,H.,: "Cloning and characterization of a rat H^+/peptide cotransporter mediating absorptoption of β-lactam antibiotics in the intestine" J.Pharmacol.Exp.ther.275(3). 1631-1637 (1995)

[文献書誌] Nagai,J.: "Specificity of p-aminohippurate transport system in the OK kidney epithelial cell line" J.Pharmacol.Exp.ther.274(3). 1161-1166 (1995)

[文献書誌] Hirai,M.: "Cepharantin,a multidrug resistant modifier,is a substrate for P-glycoprotein." J.Pharmacol.Exp.Ther.275(1). 73-78 (1995)

[文献書誌] Yoshida,A.: "Immunohistochemical localization of Na^+-dependent glucose trasnporter in the rat digestive tract." Histochmical J.27. 420-426 (1995)

[文献書誌] Nishizawa,K.: "Substrate recognition domain of the Gal2 galactose transporter in yeast Saccharomyses cerevisiae as revealed by chimeric galactose-" J.Biol.Chem.270. 2423-2426 (1995)

[文献書誌] Takigawa,T.: "Antibodies against GM1 ganglioside affect K^+ and Na^+ currents in isolated rat myelinated nerve fibers." Ann Neurology. 37. 436-442 (1995)

[文献書誌] Guo,J.: "A sustained inward current activated at the diastolic potential range in rabbit sino-atrialnode cells." J.Physiology. 483. 1-13 (1995)

[文献書誌] Manabe,K.: "Hyperpolarization induced by vasoactive substances in intact guinea-pig endocardial endothelial cells." J.Physiology. 484. 25-40 (1995)

[文献書誌] Manabe,K.: "Classification of ion channels in the luminal and abluminal membranes of guinea-pig endocardial endothelial cells." J.Physiology. 484. 41-52 (1995)

[文献書誌] Manabe,K.: "Non-selective cation current of guinea-pig endocardial endothelial cells." J.Physiology. 487. 407-419 (1995)

[文献書誌] Inagaki,N.: "Reconstitution of lKATP : An inward rectifier subunit plus the sulfonylurea receptor." Science. 270. 1166-1170 (1995)

[文献書誌] Inagaki,N.: "cDNA sequence,gene structure,and chromosomal localization of the human ATP-sensitive potassium channel,uKATP-1,Gene (KCNJ8)." Genomics. 30. 102-104 (1995)

[文献書誌] Yamada,Y.: "Human gastric inhibitory polyeptide receptor : cloning of gene and cDNA." Genomics. 29. 773-776 (1995)

[文献書誌] Kuromi,H.: "A monitor of secretion from single pancreatic β-cells." Diabetes. 44. 1213-1217 (1995)

[文献書誌] Yamada,Y.: "The structures of the human calcium channel α1 subunit (CACNL1A2) and β subnuit (CACNLB3) genes." Genomics. 27. 312-319 (1995)

[文献書誌] Marqueze,B.: "Cellular localization of Synaptotagmin I,II,and III mRNAs in the central nervous system,pituitary and adrenal glands of the rat." J.Neurosci.15. 4906-4917 (1995)

[文献書誌] Inagaki,N.: "Expression and role of ionotropic glutamate recepters in pancreatic islet cells." FASEB.J. 9. 686-691 (1995)

[文献書誌] Inagaki,N.: "Cloning and functional characterization of a novel ATP-sensitive potassium channel ubiquitously expressed in rat tissues,including" J.Biol.Chem.270. 5691-5695 (1995)

[文献書誌] Blondel,O.: "Inositol 1,4,5-trisphosphate recetpors,secretory grnules and secretion in endocrine and neuroendocrine cells." Trends Neurosci. 18. 157-161 (1995)

[文献書誌] Yorimitsu,K.: "Cloning and sequence of a human endothelin converting enzyme in renal adenocarcinoma (ACHN) cells producing endothelin-2." Biochem.Biophys.Res.Commun.208. 721-727 (1995)

[文献書誌] Jones,J.M.: "Synaptotagmin genes on mouse Chromosomes 1,7 and 10 and human Chromosome 19." Nammal.Genome. 6. 212-213 (1995)

[文献書誌] Oho,C.: "Expression and complex formation of soluble N-ethyl-maleimide-sensitive factor attachment protein (SNAP) receptors in clonal rat" Neurosci.Lett.186. 208-210 (1995)

[文献書誌] Seino,S: "CACN4,the major α1 subunit isoform of voltage-dependent calsium channels in pancreatic β-cells : α minireview of current progress." Diabetes Res.Clin.Pract.28. S99-S103 (1995)

[文献書誌] lhara,Y.: "Molecular diversity and functional characterization of voltage-dependent calcium channels (CACN4) expressed in pancreatic β-cells." Mol.Endocrinol.9. 121-130 (1995)

[文献書誌] Shin,Y.: "Reconstitution of the F_1-ATPase activty from purified α、β、γ and δ or ε subunits with glutathions S-transferase fused at their amino" Biochim.Biophys.Acta. 1273. 62-70 (1996)

[文献書誌] Sarker,R.l.: "Properties of a Na+/galactose (glucose) symport system in Vibrio parahaemolyticus." Biochim.Biophys.Acta. (in press).

[文献書誌] Sarker,R.l: "Sequence of a Na+/glucose symporter gene and its flanking regions of Vibrio parahaemolyticus." Biochim.Biophys.Acta. (in press).

[文献書誌] Kasahara,T.: "Expression of the rat GLUT1 glucose transporter in the yeast Saccharomyces cerevisiae." Biochemical.J.(in press).

[文献書誌] Ding W-G.: "A possible role of ATP-sensitive K^+ channel in determining the duration of spike-bursts in mouse pancreatic β-cells." Biochim.Biophys.Acta. (in press).

[文献書誌] He L-P.: "Glucagon induced Ca^<2+>-dependent increase of reduced pyridine nucleotides in mouse pancreatic β-cells." Biochim.Biophys.Acta. (in press).

1995 年度実績報告書

土屋友房岡山大学, 薬学部, 教授 (80012673)