Wang Lili, Zhou Hu, Li Zhengjun, Lim Teck Kwang, Lim Xin Shan, Lin Qingsong
NUS Environmental Research Institute (NERI), National University of Singapore, Singapore.
Department of Biological Sciences, National University of Singapore, Singapore.
Protein Expr Purif. 2015 Nov;115:146-52. doi: 10.1016/j.pep.2015.08.014. Epub 2015 Aug 13.
Aquaporins are integral membrane channel proteins found in all kingdoms of life. The Escherichia coli aquaporin Z (AqpZ) has been shown to solely conduct water at high permeability. Functional AqpZ is generally purified from the membrane fraction. However, the quantity of the purified protein is limited. In this study, a new method is developed to achieve high yield of bioactive AqpZ protein. A mild detergent n-dodecyl-β-D-maltopyranoside (DDM) was used to solubilize the over-expressed insoluble AqpZ from inclusion bodies without a refolding process. The recovered AqpZ protein showed high water permeability comparable with AqpZ obtained from the membrane fraction. In this way, the total yield of bioactive AqpZ has been increased greatly, which will facilitate the structural and functional characterization and future applications of AqpZ.
水通道蛋白是存在于所有生命王国中的整合膜通道蛋白。大肠杆菌水通道蛋白Z（AqpZ）已被证明在高渗透性下仅传导水。功能性AqpZ通常从膜部分纯化。然而，纯化蛋白的量是有限的。在本研究中，开发了一种新方法以实现生物活性AqpZ蛋白的高产率。使用温和的去污剂正十二烷基-β-D-麦芽糖苷（DDM）从包涵体中溶解过量表达的不溶性AqpZ，而无需重折叠过程。回收的AqpZ蛋白显示出与从膜部分获得的AqpZ相当的高水渗透性。通过这种方式，生物活性AqpZ的总产量大大提高，这将有助于AqpZ的结构和功能表征以及未来的应用。
