Hitzenberger Manuel, Zacharias Martin
Physics Department T38, Technical University of Munich, Garching, Germany.
Front Chem. 2019 Jan 4;6:640. doi: 10.3389/fchem.2018.00640. eCollection 2018.
γ-secretase, an intramembrane-cleaving aspartyl protease is involved in the cleavage of a large number of intramembrane proteins. The most prominent substrate is the amyloid precursor protein, whose proteolytic processing leads to the production of different amyloid Aβ peptides. These peptides are known to form toxic aggregates and may play a key role in Alzheimer's disease (AD). Recently, the three-dimensional structure of γ-secretase has been determined via Cryo-EM, elucidating the spatial geometry of this enzyme complex in different functional states. We have used molecular dynamics (MD) simulations to study the global dynamics and conformational transitions of γ-secretase, as well as the water and lipid distributions in and around the transmembrane domains in atomic detail. Simulations were performed on the full enzyme complex and on the membrane embedded parts alone. The simulations revealed global motions compatible with the experimental enzyme structures and indicated little dependence of the dynamics of the transmembrane domains on the soluble extracellular subunits. During the simulation on the membrane spanning part a transition between an inactive conformation (with catalytic residues far apart) toward a putatively active form (with catalytic residues in close proximity) has been observed. This conformational change is associated with a distinct rearrangement of transmembrane helices, a global compaction of the catalytically active presenilin subunit a change in the water structure near the active site and a rigidification of the protein fold. The observed conformational rearrangement allows the interpretation of the effect of several mutations on the activity of γ-secretase. A number of long-lived lipid binding sites could be identified on the membrane spanning surface of γ-secretase which may coincide with association regions of hydrophobic membrane helices to form putative substrate binding exosites.
γ-分泌酶是一种膜内裂解天冬氨酸蛋白酶,参与大量膜内蛋白的裂解。最突出的底物是淀粉样前体蛋白,其蛋白水解加工会导致产生不同的淀粉样Aβ肽。已知这些肽会形成有毒聚集体,并可能在阿尔茨海默病(AD)中起关键作用。最近,通过冷冻电镜确定了γ-分泌酶的三维结构,阐明了该酶复合物在不同功能状态下的空间几何形状。我们使用分子动力学(MD)模拟来研究γ-分泌酶的整体动力学和构象转变,以及跨膜结构域内外原子水平的水和脂质分布。对完整的酶复合物和仅对膜嵌入部分进行了模拟。模拟揭示了与实验酶结构相符的整体运动,并表明跨膜结构域的动力学对可溶性细胞外亚基的依赖性很小。在对跨膜部分的模拟过程中,观察到从无活性构象(催化残基相距很远)向假定的活性形式(催化残基紧密相邻)的转变。这种构象变化与跨膜螺旋的明显重排、催化活性早老素亚基的整体压缩、活性位点附近水结构的变化以及蛋白质折叠的刚性化有关。观察到的构象重排有助于解释几种突变对γ-分泌酶活性的影响。在γ-分泌酶的跨膜表面可以识别出许多长寿命的脂质结合位点,这些位点可能与疏水膜螺旋的结合区域重合,形成假定的底物结合外位点。