2000 年度実績報告書

血管平滑筋細胞における低分子量G蛋白rho subfamilyの役割

研究課題

研究課題/領域番号	11838013
研究機関	九州大学
研究代表者	西村淳二九州大学, 大学院・医学研究院, 助教授 (90237727)
キーワード	血管平滑筋 / 低分子量G蛋白 / カルシウム感受性 / 血管緊張 / カルシウム / ミオシン軽鎖脱リン酸化酵素
研究概要	低分子量G蛋白rho subfamily (rho A、rho B、rho C、rho G、rac 1、rac 2、rac 3、ccc42)は、細胞形態変化、細胞接着、ストレスファイバー形成、細胞遊走、細胞周期進展等、種々の細胞骨格依存性の機能に関わる重要な一群の蛋白である。近年、このrho subfamilyが、血管平滑筋の収縮や筋細胞の分化にも関与していることが明らかにされている。このrho A蛋白はその下流に存在するrho kinaseを介して、ミオシン軽鎖脱リン酸化酵素を抑制し、その結果、平滑筋のカルシウム感受性を増加させることがあきらかとなった。そこで、このミオシン軽鎖脱リン酸化酵素の2つの調節サブユニットをリコンビナント蛋白として作製し、そのスキンド標本に対する効果を検討し、昨年報告した。今年は、さらに、ミオシン軽鎖脱リン酸化酵素の調節サブユニット(M21およびM130)のゲノムの構造を明らかにし、その生理機能を明らかにした。 1、ミオシン軽鎖脱リン酸化酵素の小さい方の調節サブユニット(M21)のゲノムの構造とその生理機能。昨年報告したミオシン軽鎖脱リン酸化酵素の小さい方の調節サブユニット(M21)の内、心筋特異的に発現するHS-M21をクローニングした。このHS-M21のゲノムの構造を解析したところ、HS-M21がミオシン軽鎖脱リン酸化酵素の大きい方のサブユニットのひとつのisoformであるMYPT2と同じ遺伝子からalternative splicingにより、そのC末の部分だけが転写されることが判明した。さらにイントロン13に心筋特異的なプロモーターを見出した。HS-M21の生理機能を解析したところ、昨年報告した結果と同様に、血管平滑筋および心筋のカルシウム感受性を増加させることが判明した。さらに、このHS-M21が、ミオシン軽鎖脱リン酸化酵素の大きい方のサブユニットのひとつのisoformであるMYPT2と親和性が強く、このことがカルシウム感受性を増加させる機序と考えられた(Arimura et al., J Biol.Chem.in press)。

研究成果
(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Zhou YB: "The exogenously added small subunit of smooth muscle myosln phosphatase increases the Ca^<2+> sensitivity of the contractileapparatus in the permeabilizedporcine renal artery."Biochem Biophys Res Commun. 254. 158-163 (1999)
[文献書誌] Hirano M: "Expression, subcellular localization, and cloning of the 130-kDa regulatory subunit of myosin phosphatase in porcine aortic endothelial cells."Biochem Biophys Res Commun. 254. 490-496 (1999)
[文献書誌] Yasutsune T: "Vasorelaxationandinhibition of the voltage-operated Ca^<2+> channelsby FK506 in the porcine coronary artery."Br J Pharmacol. 126. 717-729 (1999)
[文献書誌] Zhou YB: "NH_2-terminal fragments of 130 kDa subunit of myosin phosphatase increase Ca^<2+> sensitivity of porcine renal artery."J Physiol. 516. 55-65 (1999)
[文献書誌] Kawasaki J: "Troglitazone inhibits the capacitative Ca^<2+> entry in endothelial cells."Eur J Pharmacol. 373. 111-120 (1999)
[文献書誌] Ihara E: "Thapsigaragin-inducedendothelium-dependentriphasic regulation of vascular tone in the porcine renal artery."Br J Pharmacol. 128. 689-699 (1999)
[文献書誌] Ushio-Fukai M: "Changesin the cytosolic Ca^<2+> concentrationand Ca^<2+>-sensitivity of the contractile apparatus during angiotensin II-induceddesensitization in the rabbit femoral artery."Br J Pharmacol. 129. 425-436 (2000)
[文献書誌] Ushio-Fukai M: "The mechanism of the decrease in cytosolic Ca^<2+> concentrations induced by angiotensin II in the high K^+-depolarizedrabbit femoral artery."Br J Pharmacol. 129. 437-447 (2000)
[文献書誌] Mizuno O: "Proteolysis and phosphorylation-mediated regulation of thrombin receptor activity in in situ endothelial cells."Eur J Pharmacol. 389. 13-23 (2000)
[文献書誌] Ihara E: "The mechanism of bradykinin-inducedendothelium-dependent contraction and relaxation in the porcine interlobar renal artery."Br J Pharmacol. 129. 943-952 (2000)
[文献書誌] Mizuno O: "Stimulus-specific alteration of the relationship between cytosolic Ca^<2+> transients and nitric oxide production in endothelial cells ex vivo."Br J Pharmacol. 130. 1140-1146 (2000)
[文献書誌] Kawasaki J: "Dissociation between the Ca^<2+> signal and tube formation induced by vascular endothelial growth factor in bovine aortic endothelial cells."Eur J Pharmacol. 398. 19-29 (2000)
[文献書誌] Zeng Y: "Minimal requirementfor nuclear localization of p27^<Kip1>, a cyclin-dependentkinase inhibitor."Biochem Biophys Res Commun. 274. 37-42 (2000)
[文献書誌] Shintani Y: "Mechanisms underlying the neurokininA-inducedcontraction of the pregnantrat myometrium."Br J Pharmacol. 130. 1165-1173 (2000)
[文献書誌] Kuroiwa-Matsumoto M: "Mechanisms of the thapsigargin-induced Ca^<2+> entry in in situ endothelial cells of the porcine aortic valve and the endothelium-dependentrelaxation in the porcine coronary artery."Br J Pharmacol. 131. 115-123 (2000)
[文献書誌] Hirano K: "Mitogen-induced up-regulation of non-smooth muscle isoform of α-tropomyosin in rat aortic smooth muscle cells."Eur J Pharmacol. 406. 209-218 (2000)
[文献書誌] Ieiri S: "Mechanism of vasorelaxation inducedby a novel types of Ca^<2+> channel blocker SR33805 in the porcine coronary artery."Br J Pharmacol. 131. 1592-1596 (2000)
[文献書誌] Shintani Y: "Enhanced contractile response to thrombin in the pregnant rat myometrium."Br J Pharmacol. 131. 1619-1628 (2000)
[文献書誌] Derkach DN: "Thrombin causes endothelium-dependent biphasic regulation of vascular tone in the porcine renal interlobar artery."Br J Pharmacol. 131. 1635-1642 (2000)
[文献書誌] Toshima J: "Differential effects of progesterone and 17 β-estradiol on the Ca^<2+> entry induced by thapsigargin and endothelin-1 in in situ endothelial cells."Biochem Biophys Acta. 1499. 109-12 (2000)
[文献書誌] Hirano K: "Cloning and functional expression of a degradation-resistant novel isoform of p27^<Kip1>."Biochem J. 353. 51-57 (2001)
[文献書誌] Setoguchi H: "Leukotriene C_4 enhances the contraction of porcine tracheal smooth muscle through the activation of Y-27632,a rho kinase inhibitor, sensitive pathway."Br J Pharmacol. 132. 111-118 (2001)
[文献書誌] Arimura T: "Identification, characterization and functional analysis of heart-specific myosin light chain phosphatase small subunit."J Biol Chem. (in press).
[文献書誌] Nakamura K: "Hydroxyfasudil, an active metabolite of fasudil hydrochloride, relaxes the rabbit basilar artery by disinhibition of myosin light chain phosphatase."J Cereb Blood Flow Metab. (in press).

2000 年度 実績報告書

血管平滑筋細胞における低分子量G蛋白rho subfamilyの役割

研究代表者

西村 淳二 九州大学, 大学院・医学研究院, 助教授 (90237727)

研究成果

[文献書誌] Zhou YB: "The exogenously added small subunit of smooth muscle myosln phosphatase increases the Ca^<2+> sensitivity of the contractileapparatus in the permeabilizedporcine renal artery."Biochem Biophys Res Commun. 254. 158-163 (1999)

[文献書誌] Hirano M: "Expression, subcellular localization, and cloning of the 130-kDa regulatory subunit of myosin phosphatase in porcine aortic endothelial cells."Biochem Biophys Res Commun. 254. 490-496 (1999)

[文献書誌] Yasutsune T: "Vasorelaxationandinhibition of the voltage-operated Ca^<2+> channelsby FK506 in the porcine coronary artery."Br J Pharmacol. 126. 717-729 (1999)

[文献書誌] Zhou YB: "NH_2-terminal fragments of 130 kDa subunit of myosin phosphatase increase Ca^<2+> sensitivity of porcine renal artery."J Physiol. 516. 55-65 (1999)

[文献書誌] Kawasaki J: "Troglitazone inhibits the capacitative Ca^<2+> entry in endothelial cells."Eur J Pharmacol. 373. 111-120 (1999)

[文献書誌] Ihara E: "Thapsigaragin-inducedendothelium-dependentriphasic regulation of vascular tone in the porcine renal artery."Br J Pharmacol. 128. 689-699 (1999)

[文献書誌] Ushio-Fukai M: "Changesin the cytosolic Ca^<2+> concentrationand Ca^<2+>-sensitivity of the contractile apparatus during angiotensin II-induceddesensitization in the rabbit femoral artery."Br J Pharmacol. 129. 425-436 (2000)

[文献書誌] Ushio-Fukai M: "The mechanism of the decrease in cytosolic Ca^<2+> concentrations induced by angiotensin II in the high K^+-depolarizedrabbit femoral artery."Br J Pharmacol. 129. 437-447 (2000)

[文献書誌] Mizuno O: "Proteolysis and phosphorylation-mediated regulation of thrombin receptor activity in in situ endothelial cells."Eur J Pharmacol. 389. 13-23 (2000)

[文献書誌] Ihara E: "The mechanism of bradykinin-inducedendothelium-dependent contraction and relaxation in the porcine interlobar renal artery."Br J Pharmacol. 129. 943-952 (2000)

[文献書誌] Mizuno O: "Stimulus-specific alteration of the relationship between cytosolic Ca^<2+> transients and nitric oxide production in endothelial cells ex vivo."Br J Pharmacol. 130. 1140-1146 (2000)

[文献書誌] Kawasaki J: "Dissociation between the Ca^<2+> signal and tube formation induced by vascular endothelial growth factor in bovine aortic endothelial cells."Eur J Pharmacol. 398. 19-29 (2000)

[文献書誌] Zeng Y: "Minimal requirementfor nuclear localization of p27^<Kip1>, a cyclin-dependentkinase inhibitor."Biochem Biophys Res Commun. 274. 37-42 (2000)

[文献書誌] Shintani Y: "Mechanisms underlying the neurokininA-inducedcontraction of the pregnantrat myometrium."Br J Pharmacol. 130. 1165-1173 (2000)

[文献書誌] Kuroiwa-Matsumoto M: "Mechanisms of the thapsigargin-induced Ca^<2+> entry in in situ endothelial cells of the porcine aortic valve and the endothelium-dependentrelaxation in the porcine coronary artery."Br J Pharmacol. 131. 115-123 (2000)

[文献書誌] Hirano K: "Mitogen-induced up-regulation of non-smooth muscle isoform of α-tropomyosin in rat aortic smooth muscle cells."Eur J Pharmacol. 406. 209-218 (2000)

[文献書誌] Ieiri S: "Mechanism of vasorelaxation inducedby a novel types of Ca^<2+> channel blocker SR33805 in the porcine coronary artery."Br J Pharmacol. 131. 1592-1596 (2000)

[文献書誌] Shintani Y: "Enhanced contractile response to thrombin in the pregnant rat myometrium."Br J Pharmacol. 131. 1619-1628 (2000)

[文献書誌] Derkach DN: "Thrombin causes endothelium-dependent biphasic regulation of vascular tone in the porcine renal interlobar artery."Br J Pharmacol. 131. 1635-1642 (2000)

[文献書誌] Toshima J: "Differential effects of progesterone and 17 β-estradiol on the Ca^<2+> entry induced by thapsigargin and endothelin-1 in in situ endothelial cells."Biochem Biophys Acta. 1499. 109-12 (2000)

[文献書誌] Hirano K: "Cloning and functional expression of a degradation-resistant novel isoform of p27^<Kip1>."Biochem J. 353. 51-57 (2001)

[文献書誌] Setoguchi H: "Leukotriene C_4 enhances the contraction of porcine tracheal smooth muscle through the activation of Y-27632,a rho kinase inhibitor, sensitive pathway."Br J Pharmacol. 132. 111-118 (2001)

[文献書誌] Arimura T: "Identification, characterization and functional analysis of heart-specific myosin light chain phosphatase small subunit."J Biol Chem. (in press).

[文献書誌] Nakamura K: "Hydroxyfasudil, an active metabolite of fasudil hydrochloride, relaxes the rabbit basilar artery by disinhibition of myosin light chain phosphatase."J Cereb Blood Flow Metab. (in press).

2000 年度実績報告書

西村淳二九州大学, 大学院・医学研究院, 助教授 (90237727)