1996 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, 大学院・医学研究科, 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, 大学院・医学研究科, 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 / クローニング
研究概要	イオン駆動型トランスポータとイオンチャネルについて、構造、機能、分子機構、制御機構、各系の相互協力関係等を、主として分子面から解析することを目指し、以下のような成果が得られた。 Na_+/メリビオースシンポータについて、キメラ体の作製と解析、部位特的変異導入による解析などから、イオン認識、基質認識、機能制御に関わる領域とアミノ酸残基が明らかになった。H_+/オリゴペプチドシンポータについても遺伝子クローニングと構造解析が進み、機能に重要なHis残基が同定された。 Na_+/H_+アンチポ-タについて、膜内配向性がほぼ明らかになり、また部位特異的変異導入法により機能や阻害剤の作用に重要な残基が同定された。新たに黄色ブドウ球菌で見つかったNa_+/H_+アンチポ-タはサブユニットが7つもある極めてめずらしいタイプのアンチポ-タであることがわかった。心筋のNa_+/Ca2_+アンチポ-タの特定のループ領域が自己調節能を有することがわかった。そして、この領域の部位特異的変異体の解析から、機能制御とイオン親和性に関与するアミノ酸残基が同定された。薬物トランスポータである有機アニオントランスポータの遺伝子クローニングと構造解析が行われた。酵母のグルコース輸送体とガラクトース輸送体のキメラ作製とその解析から、基質認識に最も重要なアミノ酸残基が2つ(Tyr446とTrp455)が同定された。膵β細胞のATP感受性K_+チャネルは、ABCトランスポータであるスルホニルウレア受容体(SUR)と内向き整流性K_+チャネルBIRから構成されていることが明らかにされた。膵β細胞のインスリン分泌過程におて、Na_+,K_+ATPaseとATP感受性K_+チャネルが機能的に協力しあっていることが明らかになった。

研究成果

(42件)

すべてその他

すべて文献書誌 (42件)

[文献書誌] Shin,Y.: "Reconstituon of the F1-ATPase activity from purifiedα,β,randδ or ε subunits with glutathione S-transferase fused at their amino termini." Biochim.Biophys.Acta. 1273. 62-70 (1996)
[文献書誌] Sarker,R.I.: "Properties of a Na^+ / galactose (glucose) symport system in Vibrio Parahaemolyticus." Biochim.Biophys.Acta. 1279. 149-156 (1996)
[文献書誌] Sarker,R.I.: "Sequence of a Na^+ / glucose symporter gene and its flanking regions of Vibrio parahaemolyticus." Biochim.Biophys.Acta. 1281. 1-4 (1996)
[文献書誌] Nozaki,K.: "Cloning and sequencing of the gene for Na^+ / H^+ antiporter of Vibrio parahaemolyticus." Biochem.Biophys.Res.Commun.222. 774-779 (1996)
[文献書誌] Shimamoto,T.: "Mutational analysis of the CitA citrate transporter from Salmonella typhimurium altered substrate specificity." Biohem.Biophys.Res.Commun.226. 481-487 (1996)
[文献書誌] Ohtomo,T.: "Transport of levofloxacin in a kidney epithelial cell line,LLC-PK1 Interaction with orqanic cation transporters in apical and basolatera" J.Pharmacol.Exp.Ther.276(3). 1143-1148 (1996)
[文献書誌] Ogihara,H.: "Immuno-Iocalization of H^+ / peptide cotransporter in rat digestive tract." Biochem.Biophys.Res.Commun.220(3). 848-852 (1996)
[文献書誌] Saito,H.: "Molecular cloning and tissue distribution of rat peptide transporter PEPT2." Biochim.Biophys,Acta. 1280(2). 173-177 (1996)
[文献書誌] Tanaka,K.: "Effect of cyclosporin analogues and FK506 on transcellular transport of daunorubicin and vinblastine via P-qlycoprotein." Pharm.Res.13(7). 1073-1077 (1996)
[文献書誌] Nabekura,T.: "Modulation of organic cation transport and lipid fluidity by benzyl alcohol in rat brush-border membrane." Pharm.Res.13(7). 1069-1072 (1996)
[文献書誌] Okuda,M.: "cDNA cloning and functional expression of a novel rat kidney organic cation transporter,OCT2." Biochem.Biophys.Res.Commum.224(2). 500-507 (1996)
[文献書誌] Saito,H.: "Cloning and functional characterization of a novel rat organic anion transporter mediating basolateral uptake of methotrexate in the kidney." J.Biol.Chem.271(34). 20719-20725 (1996)
[文献書誌] Takano,M.: "Regulation of p-aminohippurate transport by protein kinase C in OK kidney epithelial cells." Am.J.Physiol.271. F469-F475 (1996)
[文献書誌] Nabekura,T.: "Cholesterol modulates organic cation transport activity and lipid fludity in rat renal brush-border membranes." Biochim.Biophys.Acta. 1283(2). 232-236 (1996)
[文献書誌] Terada,T.: "Identification of the histidine residues invioved in substrate recogition by a rat H^+ / peptide cotransporter,PEPT1." FEBS Lett.394(2). 196-200 (1996)
[文献書誌] Saito,H.: "Na^+-dependent uptake of 1,5-anhydro-D-glucitol via the transport systems for D-glucose and D-mannose in kidney epithelial cell line." Jpn.J.Nephrology. 38(10). 435-440 (1996)
[文献書誌] Kasahara,T.: "Expression of the rat GLUT1 transporter in the yeast Saccharomyces cerevisiae." Biochemical J.315. 177-182 (1996)
[文献書誌] Kasahara,M.: "Transmembrane segment 10 is important for substrate recognition in Gal2 and Hxt2 sugar transporters in the veast Saccharomyces cerevisiae" FEBS Lett.389. 174-178 (1996)
[文献書誌] Iwasaki,N.: "Identification of microsatellite markers near human genes encoding the b-cell ATP-sensitive K^+ channel and linkage studies" Diabetes. 45. 267-269 (1996)
[文献書誌] Wang,C-Z.: "Cloning and pharmacological characterization of a fourn P2X receptor subtype widely expressed in brain and peripheral tissues including" Biochem.Biophys.Res.Commun.220. 196-202 (1996)
[文献書誌] Inagaki,N.: "A family of sulfonylurea feceptors determines the pharmacological properties of ATP-sensitive K^+ channels." Neuron. 16. 1011-1017 (1996)
[文献書誌] Namba,N.: "Kir2.2v : a possible negative regulator of the inwardly recifying K^+ channel Kir2.2." FEBS Lett.386. 211-214 (1996)
[文献書誌] Watabe,Y.: "Identification of histone H2A.X as a growth factor secreted by an androgen-independent subline of mouse mammary carcinoma cell." J.Biol.Chem.271. 25126-25130 (1996)
[文献書誌] Kubota,A.: "Effector coupling of somatostatin receptor subtypes on human endocrine tumors." Metabolism. 42-45 (1996)
[文献書誌] Seino,S.: "Recent progress in molecular genetic aspects of non-insulin-dependent diabetes mellitus." Internal Medicine. 35. 347-355 (1996)
[文献書誌] Seino,S.: "Molecular biology of the b-cell ATPsensitive K^+ channel." Diabetes Rev.4. 177-190 (1996)
[文献書誌] Wang,Z.: "Cell distension-induced increase of the delayed rectifier K^+ current in guinea pig ventricular myocytes." Circulation Research. 78. 466-474 (1996)
[文献書誌] Matsuoka,S.: "Multiple functional states of the cardiac Na^+-Ca^<2+> exchanger. Whole-cell,native-excised,and cloned-excised properties." Annals of the New York Academy of Sciences. 779. 159-170 (1996)
[文献書誌] Nicoll,D.A.: "Mutation of amino acid residues in the putative transmembrane segments of the cardiac sarcolemmal Na^+-Ca^<2+> exchanger." J.Biol.Chem.271. 13385-13391 (1996)
[文献書誌] Hryshko,L.V.: "Anomalous regulation of the Drosophia Na^+-Ca^<2+> exchanger by Ca^<2+>" J.General Physiology. 108. 67-74 (1996)
[文献書誌] Gou,J.: "Monovalent cation conductances of low-threshold and sustained inward current in rabit sinoatrial node cells." Pflugers Archiv. 433. 209-211 (1996)
[文献書誌] Okazaki,N.: "Characteristics of the melibiose transport and its primary strucure in Enterobacter aerogenes." Biochim.Biophys.Acta. (in press).
[文献書誌] Noumi,T.: "Identification and characterization of functional residues in a Na^+ / H^+ antiporter (NhaA) from Escherichia coli by randam mutagenesis." J.Biochem.(in press).
[文献書誌] Sarker,R.I.: "Primary structure and properties of the Na^+ / glucose symporter (SgIS) of Vibrio parahaemolvticus." J.Bacteriol.(in press).
[文献書誌] Tanaka,K.: "Relationship between expression level of P-glycoprotein and daunorubicin transport in LLC-PK1 cells trasdected with human MDR1" Biochem.Pharmacol.(in press).
[文献書誌] Yano,I.: "Evaluation of renal tubular secretion and reabsorption of levofloxacin in rats." Pharm.Res.(in press).
[文献書誌] Kasahara,T.: "Characterization of rat Glut4 glucose transporter expressed in yeast Saccharomyces cerevisiae." Biochim.Bhiophys.Acta. (in press).
[文献書誌] Takata,K.: "Transport of glucose across the blood-brain barriers." Int.Rev.Cytol.(in press).
[文献書誌] Matsuoka,S.: "Reglulation of the cardiac Na^+-Ca^<2+> exchanger by the endogenous XJP region." J.General Physiology. (in press).
[文献書誌] Dunne,M.J.: "Familial persistent hyperinsulinemic hypoglycaemia of infancy,and mutations in the sulfonylurea receptor." New Engl.J.Med.(in press).
[文献書誌] Tanaka,J.: "Cellular distribution of the P2×4 ATP receptor mRNA in the brain and peripheral organs of rats." Arch.Histol.Cytol.(in press).
[文献書誌] Yoshitomi,H.: "Involvement of MAP kinase / c-fos signaling in the inhibition of cell growth by somatostatin in insulinoma-derived cell." Am.J.Physiol.(in press).

1996 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] Shin,Y.: "Reconstituon of the F1-ATPase activity from purifiedα,β,randδ or ε subunits with glutathione S-transferase fused at their amino termini." Biochim.Biophys.Acta. 1273. 62-70 (1996)

[文献書誌] Sarker,R.I.: "Properties of a Na^+ / galactose (glucose) symport system in Vibrio Parahaemolyticus." Biochim.Biophys.Acta. 1279. 149-156 (1996)

[文献書誌] Sarker,R.I.: "Sequence of a Na^+ / glucose symporter gene and its flanking regions of Vibrio parahaemolyticus." Biochim.Biophys.Acta. 1281. 1-4 (1996)

[文献書誌] Nozaki,K.: "Cloning and sequencing of the gene for Na^+ / H^+ antiporter of Vibrio parahaemolyticus." Biochem.Biophys.Res.Commun.222. 774-779 (1996)

[文献書誌] Shimamoto,T.: "Mutational analysis of the CitA citrate transporter from Salmonella typhimurium altered substrate specificity." Biohem.Biophys.Res.Commun.226. 481-487 (1996)

[文献書誌] Ohtomo,T.: "Transport of levofloxacin in a kidney epithelial cell line,LLC-PK1 Interaction with orqanic cation transporters in apical and basolatera" J.Pharmacol.Exp.Ther.276(3). 1143-1148 (1996)

[文献書誌] Ogihara,H.: "Immuno-Iocalization of H^+ / peptide cotransporter in rat digestive tract." Biochem.Biophys.Res.Commun.220(3). 848-852 (1996)

[文献書誌] Saito,H.: "Molecular cloning and tissue distribution of rat peptide transporter PEPT2." Biochim.Biophys,Acta. 1280(2). 173-177 (1996)

[文献書誌] Tanaka,K.: "Effect of cyclosporin analogues and FK506 on transcellular transport of daunorubicin and vinblastine via P-qlycoprotein." Pharm.Res.13(7). 1073-1077 (1996)

[文献書誌] Nabekura,T.: "Modulation of organic cation transport and lipid fluidity by benzyl alcohol in rat brush-border membrane." Pharm.Res.13(7). 1069-1072 (1996)

[文献書誌] Okuda,M.: "cDNA cloning and functional expression of a novel rat kidney organic cation transporter,OCT2." Biochem.Biophys.Res.Commum.224(2). 500-507 (1996)

[文献書誌] Saito,H.: "Cloning and functional characterization of a novel rat organic anion transporter mediating basolateral uptake of methotrexate in the kidney." J.Biol.Chem.271(34). 20719-20725 (1996)

[文献書誌] Takano,M.: "Regulation of p-aminohippurate transport by protein kinase C in OK kidney epithelial cells." Am.J.Physiol.271. F469-F475 (1996)

[文献書誌] Nabekura,T.: "Cholesterol modulates organic cation transport activity and lipid fludity in rat renal brush-border membranes." Biochim.Biophys.Acta. 1283(2). 232-236 (1996)

[文献書誌] Terada,T.: "Identification of the histidine residues invioved in substrate recogition by a rat H^+ / peptide cotransporter,PEPT1." FEBS Lett.394(2). 196-200 (1996)

[文献書誌] Saito,H.: "Na^+-dependent uptake of 1,5-anhydro-D-glucitol via the transport systems for D-glucose and D-mannose in kidney epithelial cell line." Jpn.J.Nephrology. 38(10). 435-440 (1996)

[文献書誌] Kasahara,T.: "Expression of the rat GLUT1 transporter in the yeast Saccharomyces cerevisiae." Biochemical J.315. 177-182 (1996)

[文献書誌] Kasahara,M.: "Transmembrane segment 10 is important for substrate recognition in Gal2 and Hxt2 sugar transporters in the veast Saccharomyces cerevisiae" FEBS Lett.389. 174-178 (1996)

[文献書誌] Iwasaki,N.: "Identification of microsatellite markers near human genes encoding the b-cell ATP-sensitive K^+ channel and linkage studies" Diabetes. 45. 267-269 (1996)

[文献書誌] Wang,C-Z.: "Cloning and pharmacological characterization of a fourn P2X receptor subtype widely expressed in brain and peripheral tissues including" Biochem.Biophys.Res.Commun.220. 196-202 (1996)

[文献書誌] Inagaki,N.: "A family of sulfonylurea feceptors determines the pharmacological properties of ATP-sensitive K^+ channels." Neuron. 16. 1011-1017 (1996)

[文献書誌] Namba,N.: "Kir2.2v : a possible negative regulator of the inwardly recifying K^+ channel Kir2.2." FEBS Lett.386. 211-214 (1996)

[文献書誌] Watabe,Y.: "Identification of histone H2A.X as a growth factor secreted by an androgen-independent subline of mouse mammary carcinoma cell." J.Biol.Chem.271. 25126-25130 (1996)

[文献書誌] Kubota,A.: "Effector coupling of somatostatin receptor subtypes on human endocrine tumors." Metabolism. 42-45 (1996)

[文献書誌] Seino,S.: "Recent progress in molecular genetic aspects of non-insulin-dependent diabetes mellitus." Internal Medicine. 35. 347-355 (1996)

[文献書誌] Seino,S.: "Molecular biology of the b-cell ATPsensitive K^+ channel." Diabetes Rev.4. 177-190 (1996)

[文献書誌] Wang,Z.: "Cell distension-induced increase of the delayed rectifier K^+ current in guinea pig ventricular myocytes." Circulation Research. 78. 466-474 (1996)

[文献書誌] Matsuoka,S.: "Multiple functional states of the cardiac Na^+-Ca^<2+> exchanger. Whole-cell,native-excised,and cloned-excised properties." Annals of the New York Academy of Sciences. 779. 159-170 (1996)

[文献書誌] Nicoll,D.A.: "Mutation of amino acid residues in the putative transmembrane segments of the cardiac sarcolemmal Na^+-Ca^<2+> exchanger." J.Biol.Chem.271. 13385-13391 (1996)

[文献書誌] Hryshko,L.V.: "Anomalous regulation of the Drosophia Na^+-Ca^<2+> exchanger by Ca^<2+>" J.General Physiology. 108. 67-74 (1996)

[文献書誌] Gou,J.: "Monovalent cation conductances of low-threshold and sustained inward current in rabit sinoatrial node cells." Pflugers Archiv. 433. 209-211 (1996)

[文献書誌] Okazaki,N.: "Characteristics of the melibiose transport and its primary strucure in Enterobacter aerogenes." Biochim.Biophys.Acta. (in press).

[文献書誌] Noumi,T.: "Identification and characterization of functional residues in a Na^+ / H^+ antiporter (NhaA) from Escherichia coli by randam mutagenesis." J.Biochem.(in press).

[文献書誌] Sarker,R.I.: "Primary structure and properties of the Na^+ / glucose symporter (SgIS) of Vibrio parahaemolvticus." J.Bacteriol.(in press).

[文献書誌] Tanaka,K.: "Relationship between expression level of P-glycoprotein and daunorubicin transport in LLC-PK1 cells trasdected with human MDR1" Biochem.Pharmacol.(in press).

[文献書誌] Yano,I.: "Evaluation of renal tubular secretion and reabsorption of levofloxacin in rats." Pharm.Res.(in press).

[文献書誌] Kasahara,T.: "Characterization of rat Glut4 glucose transporter expressed in yeast Saccharomyces cerevisiae." Biochim.Bhiophys.Acta. (in press).

[文献書誌] Takata,K.: "Transport of glucose across the blood-brain barriers." Int.Rev.Cytol.(in press).

[文献書誌] Matsuoka,S.: "Reglulation of the cardiac Na^+-Ca^<2+> exchanger by the endogenous XJP region." J.General Physiology. (in press).

[文献書誌] Dunne,M.J.: "Familial persistent hyperinsulinemic hypoglycaemia of infancy,and mutations in the sulfonylurea receptor." New Engl.J.Med.(in press).

[文献書誌] Tanaka,J.: "Cellular distribution of the P2×4 ATP receptor mRNA in the brain and peripheral organs of rats." Arch.Histol.Cytol.(in press).

[文献書誌] Yoshitomi,H.: "Involvement of MAP kinase / c-fos signaling in the inhibition of cell growth by somatostatin in insulinoma-derived cell." Am.J.Physiol.(in press).

1996 年度実績報告書

土屋友房岡山大学, 薬学部, 教授 (80012673)