Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institute, Villigen, Switzerland.
Institute of Biochemistry, ETH Zurich, Switzerland.
PLoS One. 2018 Jun 13;13(6):e0198832. doi: 10.1371/journal.pone.0198832. eCollection 2018.
The translocator protein (TSPO) is an 18 kDa polytopic membrane protein of the outer mitochondrial membrane, abundantly present in the steroid-synthesising cells. TSPO has been linked to a number of disorders, and it is recognised as a promising drug target with a range of potential medical applications. Structural and biochemical characterisation of a mammalian TSPO requires expression and purification of the protein of high quality in sufficiently large quantities. Here we describe detailed procedures for heterologous expression and purification of mammalian TSPO in HEK293 cells. We demonstrate that the established procedures can be used for untagged TSPO as well as for C-terminally fused TSPO constructs. Our protocol can be routinely used to generate high-quality TSPO preparations for biochemical and structural studies.
转位蛋白（TSPO）是一种 18 kDa 的多跨膜蛋白，位于外线粒体膜上，在类固醇合成细胞中大量存在。TSPO 与许多疾病有关，它被认为是一种有前途的药物靶点，具有广泛的潜在医学应用。哺乳动物 TSPO 的结构和生化特性需要高质量和足够数量的蛋白质的表达和纯化。本文描述了在 HEK293 细胞中异源表达和纯化哺乳动物 TSPO 的详细程序。我们证明，所建立的程序可用于未标记的 TSPO 以及 C 端融合的 TSPO 构建体。我们的方案可常规用于生成用于生化和结构研究的高质量 TSPO 制剂。
