1995 年度実績報告書

筋線維におけるクロスブリッジ機構の分子生理学

研究課題

研究課題/領域番号	07308075
研究種目	総合研究(A)
研究機関	大分医科大学
研究代表者	山田和廣大分医科大学, 医学部, 教授 (20053027)
研究分担者	飯野正光東京大学, 医学部, 教授 (50133939) 堀田凱樹東京大学, 大学院・理学系研究科, 教授 (30010036) 杉晴夫帝京大学, 医学部, 教授 (20082076) 八木直人東北大学, 医学部, 講師 (80133940) 平塚寿章旭川医科大学, 医学部, 助教授 (30041825)
キーワード	筋肉 / 生体運動 / クロスブリッジ / 筋線維 / 蛍光プローブ / 構造変化 / X線回折 / 運動単分子
研究概要	筋肉に内在する、いわゆる生体分子モーター(クロスブリッジ)は、細胞運動から筋収縮にいたる生体運動に広く普遍的な分子的機構によって機能していると考えられている。筋肉は、生体分子モーターにおける力発生機構の研究を行うための対象として、特異な位置を占めている。クロスブリッジの力発生機構の詳細は、構造を保った系としての筋線維を対象として、はじめて明らかにされるものであると考える。本研究の目的は、クロスブリッジサイクルに存在する重要なステップについて、実際の筋線維内におけるそれぞれのクロスブリッジ状態の化学的性質、力学的性質、タンパク構造および熱力学的性質について明らかにし、それに基づいて新しいクロスブリッジ説を構築し、生体運動機構の本態に迫ることにある。山田、平塚および杉は、協力して筋線維における蛍光の変化を記録する事を試みた。その結果、新しい蛍光プローブの開発、および筋運動分子の構造変化に関する新しい情報が得られている。八木は、筋線維における構造変化のX線回折による研究をs進めた。若林は単離した運動分子の構造変化をX線回折によって追求し新しい結果を得た。片山は運動分子の構造変化を電子顕微鏡を用いて追求した。柳田は運動単分子の発生する力を記録し、その特性を詳しく調べた。児玉は、運動分子の力発生について、熱力学的理論構築を行うべく研究を進めた。飯野と大槻は、筋線維におけるカルシウム制御の分子機構を追求した。堀田は、変異筋線維の生成を目的とする研究を進めた。

研究成果
(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Emoto,Y.: "Tension relaxation induced by pulse photolysis of caged ATP in partially crosslinked fibers from rabbit psoas muscle." Proc.Natil.Acad.Sci.USA. 92. 1461-1464 (1995)
[文献書誌] Yamada,K.: "Labeling of myosin with a fluorescent analog of ATP and the fluorescent transients after photolysis of caged nucleotides in skeletal muscle fibers." Biophysical Journal. 68. 331s (1995)
[文献書誌] Kagawa,K.: "BDM compared with Pi and low Ca2+ in the cross-bridge reaction initiated by flash photolysis of caged ATP." Biophysical Journal. 68. 2590-2600 (1995)
[文献書誌] 平塚寿章: "[総説]ミオシンのATP結合ポケットの構造変化と筋収縮" 生物物理. 35. 2590-2600 (1995)
[文献書誌] Hiratsuka,T.: "Nucleotide-induced closure of the ATP-binding pocket in myosin subfragment-1" Journal of Biological Chemistry. 269. 27251-27257 (1994)
[文献書誌] Yagi,N.: "Structural changes in myosin cross-bridges during shortening of frog skeletal muscle." Journal of Muscle Research and Cell Motility. 16. 57-63 (1995)
[文献書誌] Hosoya,T.: "glial cells missing : A binary switch between neural and glial determination in Drosophila." Cell. 82. 1025-1036 (1995)
[文献書誌] Chiba,A.: "Fasciclin III as a synaptic target recognition molecule in Drosophila" Nature. 374. 166-168 (1995)
[文献書誌] Goto,S.: "Enhancer trapping of positional information corresponding to the polar coodinate system in the imaginal disc of Drosophila melanogaster." Roux's Archiv Dev.Biol.204. 378-391 (1995)
[文献書誌] Ikemoto,T.: "Enhancing effect of calmodulin on Ca2+-induced Ca2+ release in the sarcoplasmic reticulum of rabbit skeletal muscle fibres" Journal of Physiology. 487. 573-582 (1995)
[文献書誌] Takeshima,H.: "Ca2+ omdiced Ca2+ release in myocytes from dyspedic mice lacking type-1 ryanodine receptor." EMBO J.14. 1025-1036 (1995)
[文献書誌] Yamamoto,K.: "Actin based motor protein from fresh water alga Chara corallina" Journal of Molecular Biology. (in press). (1995)
[文献書誌] Katayama,E.: "Effect of caldesmon on the assamby of smooth muscle myosin." Journal of Biological Chemistry. 270(8). 3319-3925 (1995)
[文献書誌] Katayama,E.: "Mode of caldesmon binding to smooth musle thin filament : Possible projection of the amino-terminal domain of caldesmon from native thin filament." Biophysical Journal. 68(6). 2419-2428 (1995)
[文献書誌] Iwamoto,H.: "Effect of stretch and release on equatorial X-ray diffraction during a twitch contraction of frog skeletal muscle." Biophysical Journal. 68. 227-234 (1995)
[文献書誌] Sugimoto,Y.: "Conformational changes of the myosin heads during hydrolysis of ATP as analyzed by X-ray solution scattering." Biophysical Journal. 68. 29s-34s (1995)
[文献書誌] Yagi,N.: "A real-time observation of X-ray diffraction from frog skeletal muscle during and after slow length changes." Japan Journal of Physiology. 45. 583-606 (1995)
[文献書誌] Kawashima,A.: "Troponin isoform dependent pH dependence of Ca2+-activated myofibrillar ATpase activity of avian slow fast skeletal muscles." Biochem.Biophys.Res.Commun.207. 585-592 (1995)
[文献書誌] Yanagida,T.: "Force and steps generated by single myosin molecues." Biophys.J.68. 312-320 (1995)
[文献書誌] Funatsu,T.: "Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution." Nature. 374. 555-559 (1995)
[文献書誌] Higuchi,H.: "Gompliance of thin filaments in skinned fibers of rabbit skeletal muscle." Biophys.J.69. 1000-1010 (1995)
[文献書誌] Ishijima,A: "Multiple-and single-molecule analysis of the actomyosin motor using nanometerpiconewton manipulation with a microneedle : Unitary step and force." Biophys.J.70. 383-400 (1996)
[文献書誌] Vate,R.V.: "Direct observation of single kinesin molecules moving along microtubules." Nature. (in press). (1996)
[文献書誌] Sugiura,S.: "Force-velocity relations of rat cardiac myosin isozymes sliding on algal cell actin cables in vitro." BBA (Biochemica et Biophysica Acta). 1231. 69-75 (1995)
[文献書誌] Kobayashi,T.: "Effect of lateral forces on the movement of myosin-coated beads on actin cables studied using a centrifuge microscope." Japanese Journal of Physiology. 45. 177-186 (1995)
[文献書誌] Chaen,S.: "The force-velocity relationship of the ATP-dependent actin-myosin sliding causing cytoplasmic streaming in algal cells,studied using a centrifuge microscope." The Journal of Experimental Biology. 198. 1021-1027 (1995)
[文献書誌] Shirakawa,I.: "The mode of ATP-dependent microtubule-kinesin sliding in the auxotonic condition." The Journal of Experimental Biology. 198. 1809-1815 (1995)

1995 年度 実績報告書

筋線維におけるクロスブリッジ機構の分子生理学

研究代表者

山田 和廣 大分医科大学, 医学部, 教授 (20053027)

研究成果

[文献書誌] Emoto,Y.: "Tension relaxation induced by pulse photolysis of caged ATP in partially crosslinked fibers from rabbit psoas muscle." Proc.Natil.Acad.Sci.USA. 92. 1461-1464 (1995)

[文献書誌] Yamada,K.: "Labeling of myosin with a fluorescent analog of ATP and the fluorescent transients after photolysis of caged nucleotides in skeletal muscle fibers." Biophysical Journal. 68. 331s (1995)

[文献書誌] Kagawa,K.: "BDM compared with Pi and low Ca2+ in the cross-bridge reaction initiated by flash photolysis of caged ATP." Biophysical Journal. 68. 2590-2600 (1995)

[文献書誌] 平塚寿章: "[総説]ミオシンのATP結合ポケットの構造変化と筋収縮" 生物物理. 35. 2590-2600 (1995)

[文献書誌] Hiratsuka,T.: "Nucleotide-induced closure of the ATP-binding pocket in myosin subfragment-1" Journal of Biological Chemistry. 269. 27251-27257 (1994)

[文献書誌] Yagi,N.: "Structural changes in myosin cross-bridges during shortening of frog skeletal muscle." Journal of Muscle Research and Cell Motility. 16. 57-63 (1995)

[文献書誌] Hosoya,T.: "glial cells missing : A binary switch between neural and glial determination in Drosophila." Cell. 82. 1025-1036 (1995)

[文献書誌] Chiba,A.: "Fasciclin III as a synaptic target recognition molecule in Drosophila" Nature. 374. 166-168 (1995)

[文献書誌] Goto,S.: "Enhancer trapping of positional information corresponding to the polar coodinate system in the imaginal disc of Drosophila melanogaster." Roux's Archiv Dev.Biol.204. 378-391 (1995)

[文献書誌] Ikemoto,T.: "Enhancing effect of calmodulin on Ca2+-induced Ca2+ release in the sarcoplasmic reticulum of rabbit skeletal muscle fibres" Journal of Physiology. 487. 573-582 (1995)

[文献書誌] Takeshima,H.: "Ca2+ omdiced Ca2+ release in myocytes from dyspedic mice lacking type-1 ryanodine receptor." EMBO J.14. 1025-1036 (1995)

[文献書誌] Yamamoto,K.: "Actin based motor protein from fresh water alga Chara corallina" Journal of Molecular Biology. (in press). (1995)

[文献書誌] Katayama,E.: "Effect of caldesmon on the assamby of smooth muscle myosin." Journal of Biological Chemistry. 270(8). 3319-3925 (1995)

[文献書誌] Katayama,E.: "Mode of caldesmon binding to smooth musle thin filament : Possible projection of the amino-terminal domain of caldesmon from native thin filament." Biophysical Journal. 68(6). 2419-2428 (1995)

[文献書誌] Iwamoto,H.: "Effect of stretch and release on equatorial X-ray diffraction during a twitch contraction of frog skeletal muscle." Biophysical Journal. 68. 227-234 (1995)

[文献書誌] Sugimoto,Y.: "Conformational changes of the myosin heads during hydrolysis of ATP as analyzed by X-ray solution scattering." Biophysical Journal. 68. 29s-34s (1995)

[文献書誌] Yagi,N.: "A real-time observation of X-ray diffraction from frog skeletal muscle during and after slow length changes." Japan Journal of Physiology. 45. 583-606 (1995)

[文献書誌] Kawashima,A.: "Troponin isoform dependent pH dependence of Ca2+-activated myofibrillar ATpase activity of avian slow fast skeletal muscles." Biochem.Biophys.Res.Commun.207. 585-592 (1995)

[文献書誌] Yanagida,T.: "Force and steps generated by single myosin molecues." Biophys.J.68. 312-320 (1995)

[文献書誌] Funatsu,T.: "Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution." Nature. 374. 555-559 (1995)

[文献書誌] Higuchi,H.: "Gompliance of thin filaments in skinned fibers of rabbit skeletal muscle." Biophys.J.69. 1000-1010 (1995)

[文献書誌] Ishijima,A: "Multiple-and single-molecule analysis of the actomyosin motor using nanometerpiconewton manipulation with a microneedle : Unitary step and force." Biophys.J.70. 383-400 (1996)

[文献書誌] Vate,R.V.: "Direct observation of single kinesin molecules moving along microtubules." Nature. (in press). (1996)

[文献書誌] Sugiura,S.: "Force-velocity relations of rat cardiac myosin isozymes sliding on algal cell actin cables in vitro." BBA (Biochemica et Biophysica Acta). 1231. 69-75 (1995)

[文献書誌] Kobayashi,T.: "Effect of lateral forces on the movement of myosin-coated beads on actin cables studied using a centrifuge microscope." Japanese Journal of Physiology. 45. 177-186 (1995)

[文献書誌] Chaen,S.: "The force-velocity relationship of the ATP-dependent actin-myosin sliding causing cytoplasmic streaming in algal cells,studied using a centrifuge microscope." The Journal of Experimental Biology. 198. 1021-1027 (1995)

[文献書誌] Shirakawa,I.: "The mode of ATP-dependent microtubule-kinesin sliding in the auxotonic condition." The Journal of Experimental Biology. 198. 1809-1815 (1995)

1995 年度実績報告書

山田和廣大分医科大学, 医学部, 教授 (20053027)