Rhodopsin purification from dark-adapted bovine retina.
Rhodopsin purification from dark-adapted bovine retina.
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DOI：
10.1007/978-1-4939-2330-4_2
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发表时间：
2015-01-01
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影响因子：
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通讯作者：
Lodowski, David T
中科院分区：
文献类型：
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作者：
Blankenship, Elise;Lodowski, David T
视紫红质的结构和生物物理学研究长期以来依赖于肉类加工业中牛视网膜的现成可用性，以及获得此类研究所需数量和纯度的视紫红质均质制剂的相对容易性。在此，我们提出了一种模块化纯化方法，采用几种策略的组合，从牛视网膜的杆外节（ROS）的蔗糖梯度分离开始，ROS的去污剂溶解，从这种去污剂溶解的ROS开始选择性提取视紫红质，并通过尺寸排阻色谱进一步纯化，从而得到适于结晶或其它生物物理学研究的高浓度的高纯度视紫红质制剂。
Structural and biophysical studies of rhodopsin have long depended upon the ready availability of bovine retina from the meat-packing industry and the relative ease of obtaining homogenous preparations of rhodopsin in the quantities and purities necessary for such study. Herein we present a modular purification methodology employing a combination of several strategies, beginning with sucrose gradient isolation of rod outer segments (ROS) from bovine retina, detergent solubilization of ROS, selective extraction of rhodopsin starting from this detergent-solubilized ROS, and further purification via size-exclusion chromatography, resulting in a preparation of high-purity rhodopsin at high concentration suitable for crystallization or other biophysical study.