School of Chemistry , The University of Sydney , Sydney NSW 2006 , Australia.
J Am Chem Soc. 2019 Jul 10;141(27):10654-10660. doi: 10.1021/jacs.9b02116. Epub 2019 Jun 21.
Generation of chemical gradients across biological membranes of cellular compartments is a hallmark of all living systems. Here we report a proof-of-concept prototype transmembrane pumping system in liposomes. The pump uses fatty acid to fuel chloride transport, thus generating a transmembrane chloride gradient. Addition of fatty acid to phospholipid vesicles generates a transmembrane pH gradient (pH < pH), and this electrochemical H potential is harnessed by an anionophore to drive chloride efflux via H/Cl cotransport. Further addition of fatty acid efficiently fuels the system to continuously drive chloride transport against the concentration gradient, up to [Cl] 65 mM | [Cl] 100 mM, and is 1400 times more efficient than using an external fuel. Based on our findings from dissecting the H/Cl flux process with the use of different liposomal fluorescence assays, and supported by additional liposome-based C NMR and DLS studies; we proposed that the presence of an anionophore can induce asymmetric distribution of fatty acid, and contribute to another Cl flux mechanism in this system.
在细胞隔室的生物膜上生成化学梯度是所有生命系统的标志。在这里,我们报告了一个脂质体中跨膜泵送系统的概念验证原型。该泵使用脂肪酸为氯的运输提供动力,从而产生跨膜氯梯度。向磷脂囊泡中添加脂肪酸会产生跨膜 pH 梯度(pH < pH),并且这种电化学 H 势能被阴离子载体利用来通过 H/Cl 协同转运驱动氯离子流出。进一步添加脂肪酸可以有效地为系统提供动力,使其能够持续地逆浓度梯度运输氯离子,最高可达 [Cl] 65 mM | [Cl] 100 mM,并且比使用外部燃料的效率高 1400 倍。基于我们使用不同的脂质体荧光测定法对 H/Cl 通量过程进行剖析的发现,并得到了基于脂质体的 13 C NMR 和 DLS 研究的支持;我们提出阴离子载体的存在可以诱导脂肪酸的不对称分布,并为该系统中的另一种 Cl 通量机制做出贡献。