Yu Daoyong, Huang Guihong, Xu Fengxi, Wang Mengfei, Liu Shuang, Huang Fang
State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao, 266580, China,
Photosynth Res. 2014 Jun;120(3):311-21. doi: 10.1007/s11120-014-9988-5. Epub 2014 Mar 6.
Surfactants play important roles in the preparation, structural, and functional research of membrane proteins, and solubilizing and isolating membrane protein, while keeping their structural integrity and activity intact is complicated. The commercial n-Dodecyl-β-D-maltoside (DDM) and Triton X-100 (TX) were used as solubilizers to extract and purify trimeric photosystem I (PSI) complex, an important photosynthetic membrane protein complex attracting broad interests. With an optimized procedure, TX can be used as an effective surfactant to isolate and purify PSI, as a replace of the much more expensive DDM. A mechanism was proposed to interpret the solubilization process at surfactant concentrations lower than the critical solubilization concentration. PSI-TX and PSI-DDM had identical polypeptide bands, pigment compositions, oxygen consumption, and photocurrent activities. This provides an alternative procedure and paves a way for economical and large-scale trimeric PSI preparation.
表面活性剂在膜蛋白的制备、结构和功能研究以及膜蛋白的溶解和分离中发挥着重要作用,然而,在保持膜蛋白结构完整性和活性不变的同时对其进行溶解和分离却很复杂。商业用的正十二烷基-β-D-麦芽糖苷(DDM)和 Triton X-100(TX)被用作增溶剂来提取和纯化三聚体光系统 I(PSI)复合物,这是一种引起广泛关注的重要光合膜蛋白复合物。通过优化程序,TX 可以作为一种有效的表面活性剂来分离和纯化 PSI,以替代更为昂贵的 DDM。有人提出了一种机制来解释在表面活性剂浓度低于临界溶解浓度时的溶解过程。PSI-TX 和 PSI-DDM 具有相同的多肽条带、色素组成、耗氧量和光电流活性。这提供了一种替代方法,并为经济且大规模地制备三聚体 PSI 铺平了道路。