Extraction and investigation of ATPase of Tetrahymena pyriformis cilia
Burnasheva, S.A.; Faĭn, F.S.
Biokhimiia 47(2): 197-205
1982
ISSN/ISBN: 0320-9725 PMID: 6461364 Document Number: 195507
The ATPase-active proteins were solubilized from T. pyriformis of cilia by treatment of the cilia with the nonionic detergent Triton X-100. The activity of Triton-solubilized ATPase is stimulated by Ca2+ and is sensitive to ethylene glycol bis(.beta.-aminoethyl ether) N,N,N',N'-tetraacetic acid. EM has demonstrated that treatment of the cilia with Triton results in demembranation of the cilia. An electrophoretic analysis of protein composition of the cilia before and after the removal of membranes and Triton extraction has shown that dynein and tubulin, the high-MW proteins of ciliary axonemes, are not extracted into solution; the Triton extracts contained proteins with MW of 104,000, 94,000, 71,000, 62,000 and 21,000, respectively. The ATPase was purified 10.sbd.25-fold by chromatography on DEAE-Sephadex A-50. Sodium dodecylsulfate polyacrylamide gel electrophoresis revealed that the highly purified Ca2+-ATPase of cilia consists of 2 subunits with MW of 93,000 and 85,000. The role of Ca2+-ATPase in the mechanism of motility of T. pyriformis of cilia is discussed.