来自独岛菌属PRO95的视紫质是一种光驱动的钠离子泵。
Proteorhodopsin from Dokdonia sp. PRO95 is a light-driven Na+-pump.
作者信息
Bertsova Y V, Bogachev A V, Skulachev V P
机构信息
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119992, Russia.
出版信息
Biochemistry (Mosc). 2015 Apr;80(4):449-54. doi: 10.1134/S0006297915040082.
The gene encoding proteorhodopsin AEX55013 from Dokdonia sp. PRO95 was cloned and expressed in Escherichia coli cells. Illumination of the proteorhodopsin-producing E. coli cells in Na+-containing media resulted in alkalinization of the media. This response was accelerated by uncoupler CCCP and inhibited by penetrating anion SCN(-). Illumination of the cells in a sodium-free medium (made by substituting Na+ with K+) resulted in SCN(-)-stimulated and CCCP-sensitive acidification of the medium. Illumination of the proteorhodopsin-containing E. coli cells caused CCCP-resistant transmembrane sodium export from these cells. We conclude that the proteorhodopsin from the marine flavobacterium Dokdonia sp. PRO95 is a primary light-driven Na+-pump. A high level of the heterologous production in E. coli cells as well as stability and purity of the isolated protein makes this proteorhodopsin an attractive model for studying the mechanism of active sodium transmembrane translocation.
克隆了来自 Dokdonia sp. PRO95 的编码视紫质 AEX55013 的基因，并在大肠杆菌细胞中进行表达。在含 Na⁺ 的培养基中光照产生视紫质的大肠杆菌细胞会导致培养基碱化。解偶联剂 CCCP 会加速这种反应，而穿透性阴离子 SCN⁻ 会抑制该反应。在无钠培养基（用 K⁺ 替代 Na⁺ 制成）中光照细胞会导致培养基发生 SCN⁻ 刺激且对 CCCP 敏感的酸化。光照含视紫质的大肠杆菌细胞会导致这些细胞发生对 CCCP 有抗性的跨膜钠输出。我们得出结论，来自海洋黄杆菌 Dokdonia sp. PRO95 的视紫质是一种主要的光驱动 Na⁺ 泵。在大肠杆菌细胞中的高水平异源生产以及分离蛋白的稳定性和纯度使得这种视紫质成为研究活性钠跨膜转运机制的有吸引力的模型。
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