Bavi Navid, Nakayama Yoshitaka, Bavi Omid, Cox Charles D, Qin Qing-Hua, Martinac Boris
St. Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Darlinghurst, NSW 2010, Australia; Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Darlinghurst, NSW 2010, Australia;
Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Darlinghurst, NSW 2010, Australia;
Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):13864-9. doi: 10.1073/pnas.1409011111. Epub 2014 Sep 8.
The lipid bilayer plays a crucial role in gating of mechanosensitive (MS) channels. Hence it is imperative to elucidate the rheological properties of lipid membranes. Herein we introduce a framework to characterize the mechanical properties of lipid bilayers by combining micropipette aspiration (MA) with theoretical modeling. Our results reveal that excised liposome patch fluorometry is superior to traditional cell-attached MA for measuring the intrinsic mechanical properties of lipid bilayers. The computational results also indicate that unlike the uniform bilayer tension estimated by Laplace's law, bilayer tension is not uniform across the membrane patch area. Instead, the highest tension is seen at the apex of the patch and the lowest tension is encountered near the pipette wall. More importantly, there is only a negligible difference between the stress profiles of the outer and inner monolayers in the cell-attached configuration, whereas a substantial difference (∼30%) is observed in the excised configuration. Our results have far-reaching consequences for the biophysical studies of MS channels and ion channels in general, using the patch-clamp technique, and begin to unravel the difference in activity seen between MS channels in different experimental paradigms.
脂质双层在机械敏感(MS)通道的门控中起着至关重要的作用。因此,阐明脂质膜的流变学性质势在必行。在此,我们引入了一个框架,通过将微吸管抽吸(MA)与理论建模相结合来表征脂质双层的力学性质。我们的结果表明,对于测量脂质双层的固有力学性质,切除脂质体膜片荧光测定法优于传统的细胞贴附式MA。计算结果还表明,与拉普拉斯定律估计的均匀双层张力不同,双层张力在膜片区域内并不均匀。相反,在膜片顶端观察到最高张力,在吸管壁附近遇到最低张力。更重要的是,在细胞贴附配置中,外层和内层单分子层的应力分布之间只有微不足道的差异,而在切除配置中观察到显著差异(约30%)。我们的结果对于使用膜片钳技术对MS通道和一般离子通道进行生物物理研究具有深远影响,并开始揭示在不同实验范式中MS通道之间观察到活动差异的原因。