Characteristics of chemically cross-linked myosin gels.
Myosin gels, 10 mm x 10 mm x 1 mm in size, were obtained by chemical cross-linking of scallop myosin using 1-ethyl-3-(3-dimethylaminoprolyl) carbodiimide hydrochloride (EDC), glutaraldehyde (GA), or transglutaminase (TG). All myosin gels showed high Mg-ATPase activity, although it sensitively depended on the species of cross-linker used. Among cross-linkers used, myosin gel cross-linked by TG showed the highest sensitivity, almost as high as that of native myosin. The motility assay of native actin filament on the myosin gel showed that all these myosin gels can give motion to actin filament. Among them the one cross-linked by TG had the highest average velocity and the lowest threshold concentration of ATP for movement of the actin filament and the values are nearly the same as that of native myosin. In order to give the actin filament motion with preferential direction, we attempted to make myosin gel with oriented structure by applying a shear stress. Myosin gel with oriented filament array 1 cm long and 50 microm in diameter was obtained. We found that actin filaments prefer to move along the axis of the oriented myosin gel with an increased velocity.
Kakugo
Akira
A
Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan.
Sugimoto
Shin
S
Shikinaka
Kazuhiro
K
Gong
Jian Ping
JP
Osada
Yoshihito
Y
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000648:ultrastructure; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000818:Animals; D016880:Anisotropy; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D005782:Gels; D005976:Glutaral / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D008974:Mollusca; D009038:Motion; D009218:Myosins / Q000737:chemistry* / Q000648:ultrastructure; D013314:Stress, Mechanical; D011503:Transglutaminases / Q000737:chemistry
D003432:Cross-Linking Reagents; D005782:Gels; D011503:Transglutaminases; D000251:Adenosine Triphosphatases; D009218:Myosins; D005022:Ethyldimethylaminopropyl Carbodiimide; D005976:Glutaral