Krasovska Maryna V, Sefcikova Jana, Réblová Kamila, Schneider Bohdan, Walter Nils G, Sponer Jirí
Institute of Biophysics, Academy of Sciences of the Czech Republic, 61265 Brno, Czech Republic.
Biophys J. 2006 Jul 15;91(2):626-38. doi: 10.1529/biophysj.105.079368. Epub 2006 Apr 14.
The hepatitis delta virus (HDV) ribozyme is an RNA enzyme from the human pathogenic HDV. Cations play a crucial role in self-cleavage of the HDV ribozyme, by promoting both folding and chemistry. Experimental studies have revealed limited but intriguing details on the location and structural and catalytic functions of metal ions. Here, we analyze a total of approximately 200 ns of explicit-solvent molecular dynamics simulations to provide a complementary atomistic view of the binding of monovalent and divalent cations as well as water molecules to reaction precursor and product forms of the HDV ribozyme. Our simulations find that an Mg2+ cation binds stably, by both inner- and outer-sphere contacts, to the electronegative catalytic pocket of the reaction precursor, in a position to potentially support chemistry. In contrast, protonation of the catalytically involved C75 in the precursor or artificial placement of this Mg2+ into the product structure result in its swift expulsion from the active site. These findings are consistent with a concerted reaction mechanism in which C75 and hydrated Mg2+ act as general base and acid, respectively. Monovalent cations bind to the active site and elsewhere assisted by structurally bridging long-residency water molecules, but are generally delocalized.
丁型肝炎病毒(HDV)核酶是一种来自人类致病HDV的RNA酶。阳离子通过促进折叠和化学反应,在HDV核酶的自我切割中起着关键作用。实验研究揭示了关于金属离子的位置以及结构和催化功能的有限但有趣的细节。在这里,我们分析了总共约200纳秒的显式溶剂分子动力学模拟,以提供关于单价和二价阳离子以及水分子与HDV核酶反应前体和产物形式结合的互补原子视图。我们的模拟发现,一个Mg2+阳离子通过内球和外球接触稳定地结合到反应前体的带负电的催化口袋中,处于潜在支持化学反应的位置。相比之下,前体中参与催化的C75质子化或该Mg2+人工置于产物结构中会导致其迅速从活性位点排出。这些发现与一种协同反应机制一致,其中C75和水合Mg2+分别作为广义碱和酸。单价阳离子在结构上桥连长驻留水分子的协助下结合到活性位点和其他位置,但通常是离域的。