Studies of dynein from Tetrahymena cilia using agarose polyacrylamide gel electrophoresis.

ABSTRACT

Described in this report is an application of agarose-polyacrylamide gel electrophoresis, which separates protein components of crude dynein fraction (Fraction I by Gibbons) derived from Tetrahymena cilia. By this method, the fraction was separated into three protein components (designated as bands I, II and III) on the gel. When the gel was actively stained for dynein ATPase, a single band appeared, which coincided with the position of band I. A purified dynein prepared by controlled pore glass (CPG-10) column chromatography and followed by Biogel A-15m filtration showed one band on the gel at the same position as band I. These results suggest that among these three protein components, band I represents dynein and bands II and III are derived form non-ATPase protein. 'Burstic phenomenon' was also observed on their ATPase activity when axoneme or crude dynein fractions were used for ATPase assay, while the phenomenon was almost extinguished when partially purified dynein after controlled pore glass column chromatography was used as sample.