Adenosine triphosphatase in the mitotic apparatus.

摘要: Methods.-Isolation of the mitotic apparatus: The principle new method has been described in several publications, and a detailed account procedure will appear shortly.5 Sea urchin eggs (Stronglycentrotus purpuratus) are source material, providing large quantities cells synchronous division. fertilized, extranleous membranes removed, permitted to proceed into mitosis. At appropriate stage, they suspended medium consisting 1.0 Ml sucrose, 0.15 M dithiodiglycol, 0.001 EDTA. In this medium, may readily be dispersed by gentle mechanical agitation, freeing apparatus. Further washing centrifugation same removes those cytoplasmic components which not contained Solution apparatuts: outlined above isolates entire apparatus; spindles, asters, centers, chromosomes, as well yolk particles, mitochondria fat droplets normally embedded periphery asters between "rays," submicroscopic particles matrix spindles asters. short, we isolate region dividing cell, rather than particular class structures. While it is interest know chemistry operations taking place, particularly interested defining chemical properties characteristic "fibrous" system To examine this, have employed two further selective treatments following isolation purification whole (1) apparatus concentrated from sucrose-dithiodiglycol-versene then dissolved 0.53 KC1 at pH ca. 8. This isotonic original cellular should preserve mitochondria, other against osmotic lysis. fact, can observe under microscope dissolution KC1, liberating intact chromosomes sometimes remain associated metaphase anaphase groupings, central regions particles. These were removed 25,000 g or higher for 15 minutes longer, supernatant was regarded tentatively solution structural activity reduced 100,000 up 2 hours.