Center for Medical Physics and Technology, Biophysics Group, Friedrich-Alexander-University, Erlangen-Nuremberg, Erlangen, Germany.
Biochem Biophys Res Commun. 2010 Jul 30;398(3):433-7. doi: 10.1016/j.bbrc.2010.06.094. Epub 2010 Jul 1.
The focal adhesion protein vinculin has been implicated in associating with soluble and membranous phospholipids. Here, we investigated the intermolecular interactions of two vinculin tail domains with membrane phospholipids. Previous studies have shown that the tail's unstructured C-terminus affects the mechanical behavior of cells, but not the H3 region. The aim of this work was to establish whether the C-terminal or the H3 region either associate favorably with or anchor in lipid membranes. This work characterizes the energetics and dynamics of phospholipid interactions using differential scanning calorimetry (DSC) as well as circular dichroism (CD) spectroscopy. Biochemical data from tryptophan quenching and SDS-PAGE experiments support calorimetric and CD spectroscopic findings insofar that only vinculin's C-terminus inserts into lipid membranes. These in vitro results provide further insight into the mechanical behavior of vinculin tail regions in cells and contribute to the understanding of their structure and function.
粘着斑蛋白 vinculin 被认为与可溶性和膜磷脂结合。在这里，我们研究了两个 vinculin 尾部结构域与膜磷脂的分子间相互作用。以前的研究表明，尾部的无规卷曲 C 端影响细胞的机械行为，但不影响 H3 区。这项工作的目的是确定 C 端或 H3 区是有利于与脂质膜结合还是锚定在脂质膜上。这项工作使用差示扫描量热法（DSC）和圆二色性（CD）光谱法来表征磷脂相互作用的能量学和动力学。来自色氨酸猝灭和 SDS-PAGE 实验的生化数据支持量热法和 CD 光谱学的发现，即只有 vinculin 的 C 端插入脂质膜中。这些体外结果进一步深入了解了 vinculin 尾部区域在细胞中的机械行为，并有助于理解它们的结构和功能。
