Lema M J, Pluskal M G, Allen P D
Department of Anesthesia Research, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
J Chromatogr. 1989 Jun 30;491(1):61-8. doi: 10.1016/s0378-4347(00)82819-4.
High-performance ion-exchange chromatography of myosin using a DEAE-5PW packing was used to purify myosin from skeletal, cardiac and smooth muscle. This method produces high-speed resolution (30-min analysis) of myosin from contaminating myofibrillar proteins. The column has a high capacity for binding myosin (up to 1 g) and can be used for small-scale preparation of highly purified myosin. Gel analysis in the presence of sodium dodecyl sulfate showed recovery of myosin with very little contamination of other myofibrillar proteins. Myosin was also recovered from small biopsy samples (0.1 g) by a direct extraction technique with recovery of biological ATPase activity.
使用DEAE - 5PW填料的高效离子交换色谱法对肌球蛋白进行分离，可从骨骼肌、心肌和平滑肌中纯化肌球蛋白。该方法能够快速分离肌球蛋白与污染的肌原纤维蛋白（30分钟分析时间）。该色谱柱结合肌球蛋白的能力很强（高达1克），可用于小规模制备高纯度肌球蛋白。在十二烷基硫酸钠存在下进行的凝胶分析表明，回收得到的肌球蛋白几乎没有其他肌原纤维蛋白的污染。通过直接提取技术，还能从小活检样本（0.1克）中回收肌球蛋白，并保留其生物ATP酶活性。
