Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA.
Proc Natl Acad Sci U S A. 2010 Apr 20;107(16):7208-13. doi: 10.1073/pnas.0913997107. Epub 2010 Apr 5.
Cellular organelle membranes maintain their integrity, global shape, and composition despite vigorous exchange among compartments of lipids and proteins during trafficking and signaling. Organelle homeostasis involves dynamic molecular sorting mechanisms that are far from being understood. In contrast, equilibrium thermodynamics of membrane mixing and sorting, particularly the phase behavior of binary and ternary model membrane mixtures and its coupling to membrane mechanics, is relatively well characterized. Elucidating the continuous turnover of live cell membranes, however, calls for experimental and theoretical membrane models enabling manipulation and investigation of directional mass transport. Here we introduce the phenomenon of curvature-induced domain nucleation and growth in membrane mixtures with fluid phase coexistence. Membrane domains were consistently observed to nucleate precisely at the junction between a strongly curved cylindrical (tube) membrane and a pipette-aspirated giant unilamellar vesicle. This experimental geometry mimics intracellular sorting compartments, because they often show tubular-vesicular membrane regions. Nucleated domains at tube necks were observed to present diffusion barriers to the transport of lipids and proteins. We find that curvature-nucleated domains grow with characteristic parabolic time dependence that is strongly curvature-dependent. We derive an analytical model that reflects the observed growth dynamics. Numerically calculated membrane shapes furthermore allow us to elucidate mechanical details underlying curvature-dependent directed lipid transport. Our observations suggest a novel dynamic membrane sorting principle that may contribute to intracellular protein and lipid sorting and trafficking.
细胞细胞器膜在脂质和蛋白质在运输和信号转导过程中剧烈交换的情况下,仍能保持其完整性、全局形状和组成。细胞器的动态平衡涉及到分子分拣机制,这些机制远未被完全理解。相比之下,膜混合和分拣的平衡热力学,特别是二元和三元模型膜混合物的相行为及其与膜力学的耦合,相对来说已经得到了很好的描述。然而,要阐明活细胞膜的连续更新,就需要实验和理论的膜模型,以便能够操纵和研究定向质量传输。在这里,我们介绍了在具有流体相共存的膜混合物中曲率诱导的域成核和生长的现象。一致观察到膜域恰好在强弯曲的圆柱形(管状)膜和吸管抽吸的巨大单层囊泡之间的交界处成核。这种实验几何形状模拟了细胞内分拣隔室,因为它们通常显示管状-囊泡状的膜区域。在管颈处成核的域被观察到对脂质和蛋白质的运输构成扩散障碍。我们发现,曲率成核的域以强烈依赖曲率的特征抛物线时间依赖性生长。我们推导出了一个反映观察到的生长动力学的分析模型。此外,数值计算的膜形状使我们能够阐明曲率依赖性定向脂质运输的力学细节。我们的观察结果表明了一种新的动态膜分拣原理,它可能有助于细胞内蛋白质和脂质的分拣和运输。