在拴系脂质双分子层膜中重构的机械敏感型MscL离子通道的电压诱导门控。
Voltage-induced gating of the mechanosensitive MscL ion channel reconstituted in a tethered lipid bilayer membrane.
作者信息
Andersson Martin, Okeyo George, Wilson Danyell, Keizer Henk, Moe Paul, Blount Paul, Fine Daniel, Dodabalapur Ananth, Duran Randolph S
机构信息
Department of Chemistry, University of Florida, Gainesville, FL, USA.
出版信息
Biosens Bioelectron. 2008 Jan 18;23(6):919-23. doi: 10.1016/j.bios.2007.09.014. Epub 2007 Oct 3.
The mechanosensitive (MS) ion channel is gated by changes in bilayer deformation. It is functional without the presence of any other proteins and gating of the channel has been successfully achieved using conventional patch clamping techniques where a voltage has been applied together with a pressure over the membrane. Here, we have for the first time analyzed the large conducting (MscL) channel in a supported membrane using only an external electrical field. This was made possible using a newly developed technique utilizing a tethered lipid bilayer membrane (tBLM), which is part of an engineered microelectronic array chip. Single ion channel activity characteristic for MscL was obtained, albeit with lower conductivity. The ion channel was gated using solely a transmembrane potential of 300 mV. Computations demonstrate that this amount of membrane potential induces a membrane tension of 12 dyn/cm, equivalent to that calculated to gate the channel in patch clamp from pressure-induced stretching of the bilayer. These results strengthen the supposition that the MscL ion channel gates in response to stress in the lipid membrane rather than pressure across it. Furthermore, these findings illustrate the possibility of using the MscL as a release valve for engineered membrane devices; one step closer to mimicking the true function of the living cell.
机械敏感(MS)离子通道由双层膜变形的变化控制门控。它在没有任何其他蛋白质存在的情况下就能发挥功能,并且使用传统的膜片钳技术,通过在膜上同时施加电压和压力,已成功实现该通道的门控。在此,我们首次仅使用外部电场分析了支撑膜中的大电导(MscL)通道。这是通过利用一种新开发的技术实现的,该技术使用了拴系脂质双层膜(tBLM),它是一种工程微电子阵列芯片的一部分。尽管电导率较低,但仍获得了MscL特有的单离子通道活性。仅使用300 mV的跨膜电位就可控制该离子通道的门控。计算表明,这种膜电位会诱导12达因/厘米的膜张力,这与通过压力诱导双层膜拉伸在膜片钳中控制通道门控时计算得出的膜张力相当。这些结果强化了这样一种假设,即MscL离子通道是响应脂质膜中的应力而非跨膜压力来控制门控的。此外,这些发现说明了将MscL用作工程膜装置释放阀的可能性;这离模拟活细胞的真正功能又近了一步。