Non-Straub type actin from molluscan catch muscle.
We have developed a method of obtaining natural actin from smooth muscles of the bivalves on the example of the Сrenomytilus grayanus catch muscle. The muscles were previously rigorized to prevent a loss of thin filaments during homogenization and washings. Thin filaments were isolated with a low ionic strength solution in the presence of ATP and sodium pyrophosphate. Surface proteins of thin filaments-tropomyosin, troponin, calponin and some minor actin-binding proteins-were dissociated from actin filaments by increasing the ionic strength to 0.6 M KCL. Natural fibrillar actin obtained in that way depolymerizes easily in low ionic strength solutions commonly used for the extraction of Straub-type actin from acetone powder. Purification of natural actin was carried out by the polymerization-depolymerization cycle. The content of inactivated actin remaining in the supernatant is much less than at a similar purification of Straub-type actin. A comparative investigation was performed between the natural mussel actin and the Straub-type rabbit skeletal actin in terms of the key properties of actin: polymerization, activation of Mg-ATPase activity of myosin, and the electron-microscopic structure of actin polymers.
Laboratory of Cell Biophysics, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, 17 Palchevsky Str., Vladivostok 690041, Russia. Electronic address: sheludko@stl.ru.
Nikolay S
NS
Shelud'ko
Laboratory of Cell Biophysics, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, 17 Palchevsky Str., Vladivostok 690041, Russia.
Ulyana V
UV
Girich
Laboratory of Cell Biophysics, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, 17 Palchevsky Str., Vladivostok 690041, Russia.
Stanislav S
SS
Lazarev
Laboratory of Cell Biophysics, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, 17 Palchevsky Str., Vladivostok 690041, Russia.
Ilya G
IG
Vyatchin
D000199:Actins / Q000737:chemistry* / Q000302:isolation & purification* / Q000648:ultrastructure; D000818:Animals; D008970:Molecular Weight; D008974:Mollusca / Q000737:chemistry*; D009130:Muscle, Smooth / Q000737:chemistry*; D014783:Viscosity
D000199:Actins
Actin activated myosin Mg-ATPase activity; Actin polymerization and depolymerization; Molluscan catch muscle; Molluscan globular actin; Thin filament extraction