2000 Fiscal Year Annual Research Report

血管平滑筋細胞における低分子量G蛋白rho subfamilyの役割

Research Project

Project/Area Number	11838013
Research Institution	KYUSHU UNIVERSITY
Principal Investigator	西村淳二九州大学, 大学院・医学研究院, 助教授 (90237727)
Keywords	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
Research Abstract	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)。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] 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)
[Publications] 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)
[Publications] Yasutsune T: "Vasorelaxationandinhibition of the voltage-operated Ca^<2+> channelsby FK506 in the porcine coronary artery."Br J Pharmacol. 126. 717-729 (1999)
[Publications] 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)
[Publications] Kawasaki J: "Troglitazone inhibits the capacitative Ca^<2+> entry in endothelial cells."Eur J Pharmacol. 373. 111-120 (1999)
[Publications] Ihara E: "Thapsigaragin-inducedendothelium-dependentriphasic regulation of vascular tone in the porcine renal artery."Br J Pharmacol. 128. 689-699 (1999)
[Publications] 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)
[Publications] 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)
[Publications] Mizuno O: "Proteolysis and phosphorylation-mediated regulation of thrombin receptor activity in in situ endothelial cells."Eur J Pharmacol. 389. 13-23 (2000)
[Publications] Ihara E: "The mechanism of bradykinin-inducedendothelium-dependent contraction and relaxation in the porcine interlobar renal artery."Br J Pharmacol. 129. 943-952 (2000)
[Publications] 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)
[Publications] 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)
[Publications] Zeng Y: "Minimal requirementfor nuclear localization of p27^<Kip1>, a cyclin-dependentkinase inhibitor."Biochem Biophys Res Commun. 274. 37-42 (2000)
[Publications] Shintani Y: "Mechanisms underlying the neurokininA-inducedcontraction of the pregnantrat myometrium."Br J Pharmacol. 130. 1165-1173 (2000)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Shintani Y: "Enhanced contractile response to thrombin in the pregnant rat myometrium."Br J Pharmacol. 131. 1619-1628 (2000)
[Publications] Derkach DN: "Thrombin causes endothelium-dependent biphasic regulation of vascular tone in the porcine renal interlobar artery."Br J Pharmacol. 131. 1635-1642 (2000)
[Publications] 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)
[Publications] Hirano K: "Cloning and functional expression of a degradation-resistant novel isoform of p27^<Kip1>."Biochem J. 353. 51-57 (2001)
[Publications] 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)
[Publications] Arimura T: "Identification, characterization and functional analysis of heart-specific myosin light chain phosphatase small subunit."J Biol Chem. (in press).
[Publications] 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 Fiscal Year Annual Research Report

血管平滑筋細胞における低分子量G蛋白rho subfamilyの役割

Principal Investigator

西村 淳二 九州大学, 大学院・医学研究院, 助教授 (90237727)

Research Products

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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).

西村淳二九州大学, 大学院・医学研究院, 助教授 (90237727)