De Franceschi Nicola, Pezeshkian Weria, Fragasso Alessio, Bruininks Bart M H, Tsai Sean, Marrink Siewert J, Dekker Cees
Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 2629 HZDelft, The Netherlands.
Groningen Biomolecular Sciences and Biotechnology Institute and Zernike Institute for Advanced Materials, University of Groningen, Nijenborgh 7, 9747 AGGroningen, The Netherlands.
ACS Nano. 2022 Nov 28;17(2):966-78. doi: 10.1021/acsnano.2c06125.
Shape defines the structure and function of cellular membranes. In cell division, the cell membrane deforms into a "dumbbell" shape, while organelles such as the autophagosome exhibit "stomatocyte" shapes. Bottom-up in vitro reconstitution of protein machineries that stabilize or resolve the membrane necks in such deformed liposome structures is of considerable interest to characterize their function. Here we develop a DNA-nanotechnology-based approach that we call the synthetic membrane shaper (SMS), where cholesterol-linked DNA structures attach to the liposome membrane to reproducibly generate high yields of stomatocytes and dumbbells. In silico simulations confirm the shape-stabilizing role of the SMS. We show that the SMS is fully compatible with protein reconstitution by assembling bacterial divisome proteins (DynaminA, FtsZ:ZipA) at the catenoidal neck of these membrane structures. The SMS approach provides a general tool for studying protein binding to complex membrane geometries that will greatly benefit synthetic cell research.
形状决定细胞膜的结构与功能。在细胞分裂过程中,细胞膜会变形为“哑铃”状,而诸如自噬体等细胞器则呈现“口形红细胞”状。自下而上地在体外重构能够稳定或分解此类变形脂质体结构中膜颈的蛋白质机器,对于表征其功能具有相当大的意义。在此,我们开发了一种基于DNA纳米技术的方法,我们称之为合成膜塑形器(SMS),其中胆固醇连接的DNA结构附着于脂质体膜上,以可重复的方式高产率地生成口形红细胞和哑铃状结构。计算机模拟证实了SMS的形状稳定作用。我们表明,通过在这些膜结构的链状颈部组装细菌分裂体蛋白(发动蛋白A、FtsZ:ZipA),SMS与蛋白质重构完全兼容。SMS方法为研究蛋白质与复杂膜几何形状的结合提供了一个通用工具,这将极大地有益于合成细胞研究。