Departments of Biomedical Engineering, University of California, Davis, California; Chemical Engineering, University of California, Davis, California.
Chemistry, University of California, Davis, California.
Biophys J. 2018 Nov 20;115(10):1942-1955. doi: 10.1016/j.bpj.2018.09.031. Epub 2018 Oct 6.
The lipidome of plant plasma membranes-enriched in cellular phospholipids containing at least one polyunsaturated fatty acid tail and a variety of phytosterols and phytosphingolipids-is adapted to significant abiotic stresses. But how mesoscale membrane properties of these membranes such as permeability and deformability, which arise from their unique molecular compositions and corresponding lateral organization, facilitate response to global mechanical stresses is largely unknown. Here, using giant vesicles reconstituting mixtures of polyunsaturated lipids (soy phosphatidylcholine), glucosylceramide, and sitosterol common to plant membranes, we find that the membranes adopt "janus-like" domain morphologies and display anomalous solute permeabilities. The former textures the membrane with a single sterol-glucosylceramide-enriched, liquid-ordered domain separated from a liquid-disordered phase consisting primarily of soy phosphatidylcholine. When subject to osmotic downshifts, the giant unilamellar vesicles (GUVs) respond by transiently producing well-known swell-burst cycles. In each cycle, the influx of water swells the GUV, rendering the membrane tense. Subsequent rupture of the membrane through transient poration, which localizes in the liquid-disordered phase or at the domain boundaries, reduces the osmotic stress by expelling some of the excess osmolytes (and solvent) before sealing. When subject to abrupt hypertonic stress, they deform by nucleating buds at the domain phase boundaries. Remarkably, this incipient vesiculation is reversed in a statistically significant fraction of GUVs because of the interplay with solute permeation timescales, which render osmotic stresses short-lived. This, then, suggests a novel control mechanism in which an interplay of permeability and deformability regulates osmotically induced membrane deformation and limits vesiculation-induced loss of membrane material. Interestingly, recapitulation of such dynamic morphological reconfigurability-switching between budded and nonbudded morphologies-due to the interplay of membrane permeability, which temporally reverses the osmotic gradient, and domain boundaries, which select modes of deformations, might prove valuable in endowing synthetic cells with novel morphological responsiveness.
植物质膜的脂质组富含至少一个多不饱和脂肪酸尾巴和各种植物固醇和植物鞘氨醇的细胞磷脂。这些膜的质膜的介观膜性质,如通透性和变形性,源于其独特的分子组成和相应的横向组织,这些性质如何促进对全球机械压力的响应在很大程度上是未知的。在这里,我们使用重建含有多不饱和脂质(大豆卵磷脂)、葡糖神经酰胺和植物膜中常见的甾醇的混合物的巨大囊泡,发现这些膜采用“类扬尼斯”结构域形态,并表现出异常的溶质渗透性。前者用富含固醇-葡糖神经酰胺的单一液体有序域对膜进行纹理处理,与主要由大豆卵磷脂组成的液体无序相分离。当受到渗透下降时,巨大的单层囊泡(GUV)通过短暂产生众所周知的肿胀-破裂循环来响应。在每个循环中,水的流入使 GUV 膨胀,使膜紧张。随后,通过瞬时穿孔使膜破裂,这局部化在液体无序相或在域边界处,通过排出一些多余的渗透物(和溶剂)来降低渗透压,然后密封。当受到突然的高渗应激时,它们通过在域相边界处成核芽来变形。值得注意的是,由于溶质渗透时间尺度的相互作用,这种初始的囊泡化在 GUV 的很大一部分中以统计学上显著的分数被逆转,这使得渗透压短暂存在。这表明,一种新的控制机制,即渗透性和变形性的相互作用调节渗透压诱导的膜变形,并限制囊泡化诱导的膜物质损失。有趣的是,由于膜通透性的相互作用,这种动态形态重构的可再现性-在芽和非芽形态之间切换-暂时逆转渗透压梯度,以及域边界选择变形模式,可能在赋予合成细胞新的形态响应性方面具有价值。