Azuma N, Asakura A, Yagi K
J Biochem. 1975 May;77(5):973-81. doi: 10.1093/oxfordjournals.jbchem.a130823.
Actomyosin was extracted from smooth muscle of molluscan abalone with 0.1 M PPit pH 6.4. Myosin was separated from the actomyosin by centrifugation at 100,000 X g in the presence of 5 mM ATP and 10 mM MgCl2. Myosin in the supernatant was further purified by gel filtration on a Sepharose 4B column. Paramyosin contamination of the actomyosin preparation interfered with the isolation of myosin and complete removal of actin and paramyosin from the myosin has not been accomplished. The myosin appeared to consist of a single f-chain and a single g-chain, as examined by SDS-disc electrophoresis in 8 or 13.7% acrylamide gel. The ATPase [EC 3.6.1.3] activity of this myosin in 0.5 M KCL at neutral pH and at 0 degrees was rather unstable and decreased by 10-20% per day. The effects of rho-chloromercuribenzoate and EDTA on the ATPase activity were similar to those observed with other smooth muscle myosin but the dependence upon pH or KCL concentration was different.
用0.1M pH 6.4的焦磷酸哌嗪从软体动物鲍鱼的平滑肌中提取肌动球蛋白。在5mM ATP和10mM MgCl2存在的情况下，通过100,000×g离心将肌球蛋白与肌动球蛋白分离。上清液中的肌球蛋白通过在Sepharose 4B柱上进行凝胶过滤进一步纯化。肌动球蛋白制剂中的副肌球蛋白污染干扰了肌球蛋白的分离，并且尚未实现从肌球蛋白中完全去除肌动蛋白和副肌球蛋白。通过在8％或13.7％丙烯酰胺凝胶中进行SDS-圆盘电泳检查，该肌球蛋白似乎由单条f链和单条g链组成。在中性pH值和0℃下，该肌球蛋白在0.5M KCl中的ATP酶[EC 3.6.1.3]活性相当不稳定，每天下降10-20％。对氯汞苯甲酸和EDTA对ATP酶活性的影响与在其他平滑肌肌球蛋白中观察到的相似，但对pH或KCl浓度的依赖性不同。
