Institute of Biological Chemistry, Biophysics and Bioengineering, Heriot-Watt University, Edinburgh EH14 4AS, UK; Edinburgh Super-Resolution Imaging Consortium.
Department of Mathematics, Maxwell Institute, MACS, Heriot-Watt University, Edinburgh EH14 4AS, UK.
Curr Biol. 2017 Feb 6;27(3):408-414. doi: 10.1016/j.cub.2016.12.002. Epub 2017 Jan 12.
Eukaryotic plasma membrane organization theory has long been controversial, in part due to a dearth of suitably high-resolution techniques to probe molecular architecture in situ and integrate information from diverse data streams [1]. Notably, clustered patterning of membrane proteins is a commonly conserved feature across diverse protein families (reviewed in [2]), including the SNAREs [3], SM proteins [4, 5], ion channels [6, 7], and receptors (e.g., [8]). Much effort has gone into analyzing the behavior of secretory organelles [9-13], and understanding the relationship between the membrane and proximal organelles [4, 5, 12, 14] is an essential goal for cell biology as broad concepts or rules may be established. Here we explore the generally accepted model that vesicles at the plasmalemma are guided by cytoskeletal tracks to specific sites on the membrane that have clustered molecular machinery for secretion [15], organized in part by the local lipid composition [16]. To increase our understanding of these fundamental processes, we integrated nanoscopy and spectroscopy of the secretory machinery with organelle tracking data in a mathematical model, iterating with knockdown cell models. We find that repeated routes followed by successive vesicles, the re-use of similar fusion sites, and the apparently distinct vesicle "pools" are all fashioned by the Brownian behavior of organelles overlaid on navigation between non-reactive secretory protein molecular depots patterned at the plasma membrane.
真核细胞质膜组织理论长期以来一直存在争议,部分原因是缺乏适当的高分辨率技术来原位探测分子结构,并整合来自不同数据流的信息[1]。值得注意的是,膜蛋白的聚类模式是各种蛋白质家族(综述见[2])中常见的保守特征,包括 SNAREs[3]、SM 蛋白[4,5]、离子通道[6,7]和受体(例如[8])。人们已经付出了很多努力来分析分泌细胞器的行为[9-13],并且理解膜与邻近细胞器之间的关系[4,5,12,14]是细胞生物学的一个基本目标,因为可能会建立广泛的概念或规则。在这里,我们探讨了一个普遍接受的模型,即质膜上的小泡由细胞骨架轨道引导到膜上的特定位置,这些位置聚集了用于分泌的分子机制[15],部分由局部脂质组成[16]。为了增进我们对这些基本过程的理解,我们将分泌机制的纳米显微镜和光谱学与细胞器跟踪数据整合到一个数学模型中,并对敲除细胞模型进行迭代。我们发现,连续囊泡所遵循的重复路径、类似融合位点的重复使用以及明显不同的囊泡“池”,都是由细胞器在质膜上非反应性分泌蛋白分子库之间导航的布朗运动叠加而成的。