Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio.
Interdisciplinary Biophysics Graduate Program, The Ohio State University, Columbus, Ohio.
Biophys J. 2021 Apr 20;120(8):1396-1416. doi: 10.1016/j.bpj.2021.01.032. Epub 2021 Feb 9.
The VQIVYK fragment from the Tau protein, also known as PHF6, is essential for aggregation of Tau into neurofibrillary lesions associated with neurodegenerative diseases. VQIVYK itself forms amyloid fibrils composed of paired β-sheets. Therefore, the full Tau protein and VQIVYK fibrils have been intensively investigated. A central issue in these studies is polymorphism, the ability of a protein to fold into more than one structure. Using all-atom molecular simulations, we generate five stable polymorphs of VQIVYK fibrils, establish their relative free energy with umbrella sampling methods, and identify the side chain interactions that provide stability. The two most stable polymorphs, which have nearly equal free energy, are formed by interdigitation of the mostly hydrophobic VIY "face" sides of the β-sheets. Another stable polymorph is formed by interdigitation of the QVK "back" sides. When we turn to examine structures from cryo-electron microscopy experiments on Tau filaments taken from diseased patients or generated in vitro, we find that the pattern of side chain interactions found in the two most stable face-to-face as well as the back-to-back polymorphs are recapitulated in amyloid structures of the full protein. Thus, our studies suggest that the interactions stabilizing PHF6 fibrils explain the amyloidogenicity of the VQIVYK motif within the full Tau protein and provide justification for the use of VQIVYK fibrils as a test bed for the design of molecules that identify or inhibit amyloid structures.
Tau 蛋白的 VQIVYK 片段,也称为 PHF6,对于 Tau 蛋白聚集形成与神经退行性疾病相关的神经纤维缠结是必不可少的。VQIVYK 自身形成由配对 β-折叠组成的淀粉样纤维。因此,全长 Tau 蛋白和 VQIVYK 纤维已被广泛研究。这些研究的一个核心问题是多态性,即蛋白质折叠成一种以上结构的能力。我们使用全原子分子模拟生成了 VQIVYK 纤维的五个稳定多态体,使用伞形采样方法确定它们的相对自由能,并确定提供稳定性的侧链相互作用。两种最稳定的多态体,其自由能几乎相等,是由β-折叠的主要疏水性 VIY“面”侧的相互交错形成的。另一种稳定的多态体是由 QVK“背面”的相互交错形成的。当我们转而检查来自患病患者或在体外生成的 Tau 纤维的低温电子显微镜实验的结构时,我们发现,在两种最稳定的面对面以及背对背多态体中发现的侧链相互作用模式在全长蛋白质的淀粉样结构中得到了重现。因此,我们的研究表明,稳定 PHF6 纤维的相互作用解释了全长 Tau 蛋白中 VQIVYK 基序的淀粉样形成性,并为使用 VQIVYK 纤维作为鉴定或抑制淀粉样结构的分子设计的试验台提供了依据。