| Project/Area Number |
01480249
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Ehime University |
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Principal Investigator |
HIWADA Kunio Ehime University School of Medicine, Professor, 医学部, 教授 (00108391)
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| Co-Investigator(Kenkyū-buntansha) |
SHIGEMATSU Yuuji Ehime University School of Medicine, Associate, 医学部附属病院, 助手 (90206087)
MURAKAMI Eiki Ehime University School of Medicine, Ex-Associate Professor, 医学部, 助教授 (90110832)
SEKIYA Michihito Ehime University Hospital, Lecturer, 医学部附属病院, 講師 (50171343)
KITAMI Yutaka Ehime University School of Medicine, Associate, 医学部, 助手 (10234270)
HAMADA Mareomi Ehime University School of Medicine, Associate Professor, 医学部, 助教授 (30127906)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1991: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1990: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1989: ¥3,300,000 (Direct Cost: ¥3,300,000)
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| Keywords | calponin / thin filament / actomyosin MgATPase / SM22 / SM22cDNA cloning / calponin cDNA cloning / chicken gizzard / rat aorta / 血管平滑筋 / cDNAクロ-ニング / ラット大動脈 / 細い線維 / 平滑筋 / ミオシン軽鎖MgATPase / cDNA / SM22α / ミオシン軽鎖MgATPase活性 / ウシ大動脈 |
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| Research Abstract |
Calponin is actin-, tropomyosin binding protein which we found in the smooth muscle cells. In this project we studied the localization of calponin in smooth muscle cell and the physiological function of calponin.
(1) We obtained thin filaments which contained actin, tropomyosin, caldesmon and calponin in molar rations of 7 : 0.9 : 0.6 : 0.7. Co-localization of actin and calponin was shown along stress fibers of smooth muscle cells from rat aorta.
(2) We studied the function of calponin in vitro. Calponin inhibited the actinactivated myosin MgATPase activity in a dose-dependent manner. This inhibition was Ca-independent. The decrease in enzymatic activity of myosin was correlated with binding of calponin to actin-tropgmypsin filaments. Caldesmon showed a further inhibition of the calponin-induced inhibition of MgATPase activity of the thiophosphorylated myosin. Calponin-induced inhibition of the myosin MgATPase was reversed by the addition of calmodulin in the presence of Ca. These results suggest that calponin acts as an inhibitory component of smooth muscle thin filaments.
(3) We performed cDNA cloning of SM22 from the chicken gizzard smooth muscle and rat aorta smooth muscle. Cloned cDNA encoding SM22 from the chicken gizzard smooth muscle had a total length of 1214bp and contained a single open reading frame which encodes 200 amino acids with a calculated molecular weight 22214. The predicted amino acid sequence was in complete agreement with the sequence determined by Pearlstone et al. who discovered the protein, except for two additional residues, isoleucine and serine at the C-terminus. The cDNA from rat aorta was 1186bp. in length and encoded 201 amino acids (Mr 22602). The predicted amino acid sequence showed an 85% Tdentity with the chicken gizzard SM22.
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