Purification of VDAC1 from rat liver mitochondria.
Purification of VDAC1 from rat liver mitochondria.
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DOI：
10.1101/pdb.prot073130
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发表时间：
2014-01-01
影响因子：
--
通讯作者：
Shoshan-Barmatz, Varda
中科院分区：
文献类型：
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作者：
Ben-Hail, Danya;Shoshan-Barmatz, Varda
为了对线粒体电压依赖性阴离子选择性通道蛋白 1 (VDAC1) 的成孔活性等功能进行生物物理测量，首先需要获得纯化的 VDAC 蛋白来源。在此协议中，我们提出了一种获得大鼠肝线粒体作为 VDAC1 来源的方法，然后描述了两种方法，一种使用非离子洗涤剂，另一种使用离子洗涤剂，用于从分离的线粒体中纯化 VDAC1。这产生了适合随后掺入人工制备的磷脂双层的 VDAC1 蛋白来源。此外，分离的线粒体可用于测定线粒体通透性转换孔（MPTP）。
To make biophysical measurements of functions such as the pore-forming activity of mitochondrial voltage-dependent anion-selective channel protein 1 (VDAC1), it is first necessary to obtain a source of purified VDAC protein. In this protocol, we present a method for obtaining rat liver mitochondria as a source of VDAC1 and then describe two methods, one using a nonionic detergent and the other an ionic detergent, for purifying VDAC1 from the isolated mitochondria. This produces a source of VDAC1 proteins that are suitable for subsequent incorporation into artificially prepared phospholipid bilayers. Furthermore, the isolated mitochondria can be used for assaying the mitochondrial permeability transition pore (MPTP).