Department of Biological and Medical Sciences, Oxford Brookes University, Oxford OX3 0BP, United Kingdom.
Proc Natl Acad Sci U S A. 2012 Jul 31;109(31):12805-10. doi: 10.1073/pnas.1202040109. Epub 2012 Jun 11.
A cell membrane can be considered a liquid-phase plane in which lipids and proteins theoretically are free to diffuse. Numerous reports, however, describe retarded diffusion of membrane proteins in animal cells. This anomalous diffusion results from a combination of structuring factors including protein-protein interactions, cytoskeleton corralling, and lipid organization into microdomains. In plant cells, plasma-membrane (PM) proteins have been described as relatively immobile, but the control mechanisms that structure the PM have not been studied. Here, we use fluorescence recovery after photobleaching to estimate mobility of a set of minimal PM proteins. These proteins consist only of a PM-anchoring domain fused to a fluorescent protein, but their mobilities remained limited, as is the case for many full-length proteins. Neither the cytoskeleton nor membrane microdomain structure was involved in constraining the diffusion of these proteins. The cell wall, however, was shown to have a crucial role in immobilizing PM proteins. In addition, by single-molecule fluorescence imaging we confirmed that the pattern of cellulose deposition in the cell wall affects the trajectory and speed of PM protein diffusion. Regulation of PM protein dynamics by the plant cell wall can be interpreted as a mechanism for regulating protein interactions in processes such as trafficking and signal transduction.
细胞膜可以被视为一个液相平面,其中脂质和蛋白质理论上可以自由扩散。然而,许多报道描述了动物细胞中膜蛋白扩散的减缓。这种异常扩散是由结构因素的组合引起的,包括蛋白质-蛋白质相互作用、细胞骨架的牵制和脂质组织成微区。在植物细胞中,质膜(PM)蛋白被描述为相对不活跃,但尚未研究构成 PM 的结构控制机制。在这里,我们使用光漂白后荧光恢复来估计一组最小 PM 蛋白的流动性。这些蛋白质仅由一个质膜锚定域融合到荧光蛋白组成,但它们的流动性仍然受到限制,就像许多全长蛋白一样。细胞骨架和膜微区结构都没有参与限制这些蛋白质的扩散。然而,细胞壁被证明在固定 PM 蛋白方面起着关键作用。此外,通过单分子荧光成像,我们证实细胞壁中纤维素沉积的模式会影响 PM 蛋白扩散的轨迹和速度。植物细胞壁对 PM 蛋白动力学的调节可以解释为一种调节运输和信号转导等过程中蛋白质相互作用的机制。