Myosin and adenosinetriphosphatase

作者：Kenneth Bailey · 发表于：Biochemical Journal · 年份：1942 · DOI：10.1042/bj0360121 · 被引用次数：312 · 研究领域：Enzyme Structure and Function、Photosynthetic Processes and Mechanisms、ATP Synthase and ATPases Research

Preparation and properties of myosinPreparation of various myosins.The tissues used were the hind-leg and longissimus dorsi muscles of the rabbit, the pectoral muscle of chicken, the leg muscle,of the horse, the hind-leg muscle of the toad and frog, and the trimmed heart muscle of the pig and rabbit.The rabbits and chickens were killed by stunning and bled via the inferior vena cava; frogs and toads were pithed after leaving the animals in the cold room at 00.The muscle of those animals killed in the laboratory could be excised and minced within 10 min.after death, but material from the slaughterhouse was usually obtained about an hour after killing.In one instance, however, a sample of horse muscle was acquired within 15 min.after bleeding.Immediately after mincing, the tissue was added to 5 vol. of ice-cold salt solution containing KCI (0.5 M) and NaHCO3 (0.03 N).This extractant has been adopted by Greenstein & Edsall [1940] in an improved method for the preparation of myosin.Solid NaHCO3 was now added at intervals to maintain the pH at 7-7 5, and after stirring slowly for an hour at 00, the stroma was centrifuged down.The opalescent extract was filtered clear through a 2 in.layer of paper pulp (saturated with extractant) and diluted with 20 vol. of ice-cold water.Dilute HCI was now added to adjust the pH of the medium to 6-8-7-0, and after settling, the supernatant liquor was decanted and the myosin centri- fuged down.For purification it was redissolved by the addition of ...