1990 Fiscal Year Annual Research Report

滑り運動系の微細構造

Research Project

Project/Area Number	02239105
Research Institution	Okazaki National Research Institutes
Principal Investigator	月田承一郎岡崎国立共同研究機構, 生理学研究所, 教授 (50155347)
Co-Investigator(Kenkyū-buntansha)	若林克三大阪大学, 基礎工学部, 助手 (00029521) 盛田フミ北海道大学, 理学部, 教授 (80000818) 八木直人東北大学, 医学部, 助手 (80133940) 沼田治筑波大学, 生物科学系, 講師 (50189354) 岡本洋順天堂大学, 医学部, 講師 (70103877)
Keywords	滑り運動 / アクチン / ミオシン / 電子顕微鏡 / 急速凍結 / X線回折
Research Abstract	昨年度の成果を踏まえて,アクチンミオシン系の滑り運動系の動的微細構造の解析を進めた。本年度の本研究班の成果のうち,最も注目すべきものは,Caged化合物を用いた急速凍結電子顕微鏡法の完成と,その応用である。以下に本年度の成果をまとめる。 1.Caged化合物を用いた急速凍結電子顕微鏡法の開発(月田承一郎):昨年度より開発をはじめていたが,本年度はすべての技術的問題点を克服し,ほぼ完成した。この凍結システムを用いて2つの実験を行ないつつある。第一は,Caged ATPを利用して、アクトS1複合体から,S1が解離していく様子を,ミリ秒時間分解能のコマ取り電子顕微鏡像として追跡していこうとするものである。多くの努力の結果,S1がアクチンフィラメントからその結合様式を変えながら離れていく様子をとらえることに成功した。その像の解釈は,これから慎重に行なわれなくてはならないが,滑り運動の分子機構を考える上で,極めて貴重なデ-タが得られたものと思われる。第二の実験は,やはりCaged ATPを利用して,本当に最大収縮速度で収縮(短縮)している途中のミオシンクロスブリッジの姿をとらえようとするものである。現在,そのような姿がとらえはじめており,最終年度の解析が期待できる。 2.ミオシン分子上のアクチン結合部位が詳しく解析された(山本・盛田) 3.酵母を用いて,種々の遺伝子組換えアクチンを作製する道を初めて開いた(沼田治)。 4.収縮(短縮)中の骨格筋からX線回折パタ-ンを得ることに成功しつつある(八木直人・若林克三)。 5.月田らとは異なった方向から,Caged化合物と電顕像を結びつける試みを行ない,見通しがたつようになった(徳永万喜洋)。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Tsukita,Sh.: "The undercoat of adherens junctions:A key specialized structure in organogenesis and carcinogenesis." Cell Struct.Funct.15. 7-12 (1990)
[Publications] Nishikawa,S.: "Localization of adherens junction proteins along the possible sliding interface between secretory ameloblasts of the rat incisor." Cell Struct.Funct.15. 245-250 (1990)
[Publications] Sato,N.: "Radixin,a barbedーendーcapping actinーmodulating protein,is concentrated at the cleavage furrow during cytokinesis." J.Cell Biol.
[Publications] Muto,E.: "Doubleーrowed organization of inner dynein arms in Chlamydomonas flagella revealed by tiltーseries thinーsection electron microscopy." J.Cell Sci.
[Publications] Tsukita,Sa.: "Specific protoーoncogenic tyrosine kinases of src family are enriched in cellーtoーcell adherens junctions where the level of tyrosine phosphorylation is elevated." J.Cell Biol.
[Publications] Tokunaga,M: "Structure and structural changes of the myosin head." Advances in Biophysics.
[Publications] Yamamoto,K.: "Shift of binding site at the interface between actin and myosin." Biochemistry. 29. 844-848 (1990)
[Publications] Yamamoto,K.: "Identification of the site important for the actinーactivated MgATPase activity of myosin subfragmentー1." J.Mol.Biol.217. 229-233 (1991)
[Publications] Hirono,M.: "“Tetrahymena actin":Copolymerization with skeletal muscle actin and interactions with muscle actinーbinding proteins." J.Biochem.107. 32-36 (1990)
[Publications] Edamatsu,M.: "Purification and characterization of Tetrahymena profilin." B.B.R.C.170. 957-962 (1990)
[Publications] Matsubara,I.: "Crossーbridge movement in fast and slow skeletal muscles of the chick." J.Physiol.
[Publications] Suzuki,R.: "Inhibition of actomyosin subfragment 1 ATPase activity by analog peptide of the actinーbinding site around the Cys(SH1) of myosin heavy chain" J.Biol.Chem.265. 4939-4943 (1990)
[Publications] Inoue,K.: "Caーdependent protein phosphatase which dephosphorylates regulatory light chainーa in scallop smooth muscle myosin." J.Biochem.107. 872-878 (1990)
[Publications] Eto,M.: "Roles of the amino acid side chains in the actinーbinding Sーsite of myosin heavy chain." J.Biochem.108. 499-504 (1990)
[Publications] Hasegawa,Y.: "Regulatory muchanism by the phosphorylation of 20ーkDa light chain of porcine aorta smooth muscle myosin." J.Biochem.108. 909-913 (1990)
[Publications] Tregear,R.T.: "Xーray diffraction and electron microscopy from Lethocerus flight muscle partially relaxed by Adenylylimidodiphosphate and ethylene glycol." J.Mol.Biol.214. 129-141 (1990)
[Publications] Konishi,M.: "Timeーresolved Xーray differaction studies of a single frog skeletal fiber." Biophys.Chem.
[Publications] Wakabayashi,K.: "Progress in Xーray synchrotron diffraction studies of muscle contraction." Synchrotron Radiation Handbook.

1990 Fiscal Year Annual Research Report

滑り運動系の微細構造

Principal Investigator

月田 承一郎 岡崎国立共同研究機構, 生理学研究所, 教授 (50155347)

Research Products

[Publications] Tsukita,Sh.: "The undercoat of adherens junctions:A key specialized structure in organogenesis and carcinogenesis." Cell Struct.Funct.15. 7-12 (1990)

[Publications] Nishikawa,S.: "Localization of adherens junction proteins along the possible sliding interface between secretory ameloblasts of the rat incisor." Cell Struct.Funct.15. 245-250 (1990)

[Publications] Sato,N.: "Radixin,a barbedーendーcapping actinーmodulating protein,is concentrated at the cleavage furrow during cytokinesis." J.Cell Biol.

[Publications] Muto,E.: "Doubleーrowed organization of inner dynein arms in Chlamydomonas flagella revealed by tiltーseries thinーsection electron microscopy." J.Cell Sci.

[Publications] Tsukita,Sa.: "Specific protoーoncogenic tyrosine kinases of src family are enriched in cellーtoーcell adherens junctions where the level of tyrosine phosphorylation is elevated." J.Cell Biol.

[Publications] Tokunaga,M: "Structure and structural changes of the myosin head." Advances in Biophysics.

[Publications] Yamamoto,K.: "Shift of binding site at the interface between actin and myosin." Biochemistry. 29. 844-848 (1990)

[Publications] Yamamoto,K.: "Identification of the site important for the actinーactivated MgATPase activity of myosin subfragmentー1." J.Mol.Biol.217. 229-233 (1991)

[Publications] Hirono,M.: "“Tetrahymena actin":Copolymerization with skeletal muscle actin and interactions with muscle actinーbinding proteins." J.Biochem.107. 32-36 (1990)

[Publications] Edamatsu,M.: "Purification and characterization of Tetrahymena profilin." B.B.R.C.170. 957-962 (1990)

[Publications] Matsubara,I.: "Crossーbridge movement in fast and slow skeletal muscles of the chick." J.Physiol.

[Publications] Suzuki,R.: "Inhibition of actomyosin subfragment 1 ATPase activity by analog peptide of the actinーbinding site around the Cys(SH1) of myosin heavy chain" J.Biol.Chem.265. 4939-4943 (1990)

[Publications] Inoue,K.: "Caーdependent protein phosphatase which dephosphorylates regulatory light chainーa in scallop smooth muscle myosin." J.Biochem.107. 872-878 (1990)

[Publications] Eto,M.: "Roles of the amino acid side chains in the actinーbinding Sーsite of myosin heavy chain." J.Biochem.108. 499-504 (1990)

[Publications] Hasegawa,Y.: "Regulatory muchanism by the phosphorylation of 20ーkDa light chain of porcine aorta smooth muscle myosin." J.Biochem.108. 909-913 (1990)

[Publications] Tregear,R.T.: "Xーray diffraction and electron microscopy from Lethocerus flight muscle partially relaxed by Adenylylimidodiphosphate and ethylene glycol." J.Mol.Biol.214. 129-141 (1990)

[Publications] Konishi,M.: "Timeーresolved Xーray differaction studies of a single frog skeletal fiber." Biophys.Chem.

[Publications] Wakabayashi,K.: "Progress in Xーray synchrotron diffraction studies of muscle contraction." Synchrotron Radiation Handbook.

月田承一郎岡崎国立共同研究機構, 生理学研究所, 教授 (50155347)