从纯化的牛心细胞色素 bc1 中去除结合的 Triton X-100。
Removal of bound Triton X-100 from purified bovine heart cytochrome bc1.
机构信息
Department of Biochemistry, University of Texas Health Science Center, San Antonio, TX 78229, USA.
出版信息
Anal Biochem. 2009 Dec 15;395(2):268-70. doi: 10.1016/j.ab.2009.08.042. Epub 2009 Sep 3.
Abstract
Cytochrome bc(1) isolated from Triton X-100-solubilized mitochondrial membranes contains up to 120 nmol of Triton X-100 bound per nanomole of the enzyme. Purified cytochrome bc(1) is fully active; however, protein-bound Triton X-100 significantly interferes with structural studies of the enzyme. Removal of Triton X-100 bound to bovine cytochrome bc(1) was accomplished by incubation with Bio-Beads SM-2 in the presence of sodium cholate. Sodium cholate is critical because it does not interfere with the adsorption of protein on the hydrophobic surface of the beads. The resulting Triton X-100-free cytochrome bc(1) retained nearly full activity, absorption spectra, subunit, and phospholipid composition.
摘要
从 Triton X-100 溶解的线粒体膜中分离出的细胞色素 bc(1) 每纳摩尔酶结合多达 120 毫摩尔 Triton X-100。纯化的细胞色素 bc(1) 完全具有活性；然而，蛋白质结合的 Triton X-100 会严重干扰酶的结构研究。通过在存在胆酸钠的情况下与 Bio-Beads SM-2 孵育，从牛细胞色素 bc(1) 上去除结合的 Triton X-100。胆酸钠是关键的，因为它不会干扰蛋白质在珠状疏水表面上的吸附。得到的无 Triton X-100 的细胞色素 bc(1) 保留了几乎全部的活性、吸收光谱、亚基和磷脂组成。
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