Arnold Alexandre, Paris Michaël, Auger Michèle
Département de Chimie, Centre de Recherche en Sciences et Ingénierie des Macromolécules, Université Laval, Québec, Canada.
Biophys J. 2004 Oct;87(4):2456-69. doi: 10.1529/biophysj.104.043729.
Lateral diffusion is an essential process for the functioning of biological membranes. Solid-state nuclear magnetic resonance (NMR) is, a priori, a well-suited technique to study lateral diffusion within a heterogeneous environment such as the cell membrane. Moreover, restriction of lateral motions by lateral heterogeneities can be used as a means to characterize their geometry. The goal of this work is to understand the advantages and limitations of solid-state NMR exchange experiments in the study of obstructed lateral diffusion in model membranes. For this purpose, simulations of lateral diffusion on a sphere with varying numbers and sizes of immobile obstacles and different percolation properties were performed. From the results of these simulations, two-dimensional 31P NMR exchange maps and time-dependent autocorrelation functions were calculated. The results indicate that the technique is highly sensitive to percolation properties, total obstacle area, and, within certain limits, obstacle size. A practical example is shown, namely the study of the well-characterized DMPC-DSPC binary mixture. The comparison of experimental and simulated results yielded obstacle sizes in the range of hundreds of nanometers, therefore bridging the gap between previously published NMR and fluorescence recovery after photobleaching results. The method could also be applied to the study of membrane protein lateral diffusion in model membranes.
横向扩散是生物膜发挥功能的一个重要过程。从先验角度来看,固态核磁共振(NMR)是一种非常适合用于研究诸如细胞膜这种异质环境中横向扩散的技术。此外,横向异质性对横向运动的限制可被用作表征其几何形状的一种手段。这项工作的目标是了解固态NMR交换实验在研究模型膜中受阻横向扩散时的优点和局限性。为此,对具有不同数量、大小的固定障碍物以及不同渗流特性的球体上的横向扩散进行了模拟。根据这些模拟结果,计算了二维³¹P NMR交换图谱和随时间变化的自相关函数。结果表明,该技术对渗流特性、总障碍物面积以及在一定限度内的障碍物大小高度敏感。给出了一个实际例子,即对特征明确的二肉豆蔻酰磷脂酰胆碱 - 二硬脂酰磷脂酰胆碱二元混合物的研究。实验结果与模拟结果的比较得出障碍物大小在数百纳米范围内,从而弥合了先前发表的NMR结果与光漂白后荧光恢复结果之间的差距。该方法也可应用于研究模型膜中膜蛋白的横向扩散。