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3. The effect of calcium on the movement of actin filaments on the mutant full length myosin-coated glass surface.
4. The effect of calcium on the sliding velocity of actin-filaments on a glass surface coated with mutant HMM.
考察
To understand regulation of myosin II isoforms by Ca2+ comprehensively, i. e., including both activation and inhibition modes, we have expressed the hybrid HMM, it is consisted of physarum heavy chain, physarum regulatory light chain and scallop essential light chain, expecting the functional hybrid HMM. But we found that scallop essential light chain could not bind to physarum heavy chain.
参考文献
1. Kohama K, Kohama T, Kendrick-Jones J. The inhibitory Ca2+-regulation of the actin-activaed Mg-ATPase activity of myosin from Physarum polycepharum plasmodia. J Biochem. 1986, 99, 1433-1446.
2. Hozumi Kawamichi. Characterization of recombinant Heavy Meromyosin of Physarum polycephalum. Kitakanto Med J. 2002, 52, 89-97.
3. Kohama K, Sohda M, Maruyama K, et al. Domain structure of Physarum myosin heavy chain. Protoplasma. 1988, 2, 37-47.
4. Kohama K, Kohno T, Okagaki T, et al. Role of actin in the myosin-linked Ca2+-regulation of ATP-dependent interaction between actin and myosin of a lower eukayote, Physarum polycephalum. J Biochem. 1991, 110, 508-513.
5. Akio Nakamura and Kohama,K. Calcium regulation of the actin-myosin interaction of Physarum polycephalum. International Review of Cytology, 1999, 191, 53-98.
6. Ogihara, S., Ikebe, M., Takahashi, K. and Tonomura, Y. Requirement of phosphorylation of Physarum myosin heavy chain for thick filament formation, actin activation of Mg-ATPase activity, and Ca2+-inhibitory superprecipitation. J Biochem. 1983, 93, 205-223.
7. Farkas, L., Andra' s Ma' lna' si-Csizmadia, Nakamura, A., Kohama, K. and La' szlo' Nyitray. Localization and characterization of the inhibitory Ca2+-binding site of Physarum polycephalum Myosin II. The Journal of Biological Chemistry. 2003, 278, 27399-27405.
8. Kohama K. Heterogeneity of amino acid incorporation rate in adult skeletal muscle actin. J Biochem. 1980, 87, 997-999.
注：
（1）本研究は2007年3月16日に日本薬理会で「カルシウム感受性真性粘菌ミオシンIIの発現と性質」発表.
（2）Hozumi Kawamichi, Ying Zhang, Mizuki Hino, Akio Nakamura, Hideyuki Tanaka, Lászlo Farkas, Lászlo Nyitray, Kazuhiro Kohama. Calcium inhibition of Physarum myosin as examined by the recombinant heavy mero-myosin. In: Regulatory mechanisms of striated muscle contraction. Eds: S.Ebashi & I. Ohtsuki. pp257-264, Springer Verlag, (2006)
作成日 2007/03/01
1. SDS-PAGE of purification steps of recombinant myosin of Physarum polycephalum
1: Molecular weight marker
2: Sup. of Sf-9 unint'ected homogenate
3: Ppt. of Sf-9 homogenate infected with both HC and LCs
4: Sup. of Sf-9 homogenate infected with both HC and LCs.
5: Sup. after the ultracentrifugation in the presence of ATP
6: effluence after using Ni-NTA column
7-10: Purified physarum myosin by using Ni-NTA column
11-15: Purified physarum myosin by using SuperoseTM6 column with HPLC
2. SDS-PAGE of purified recombinant HMM of Physarum polycephalum
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