Laboratory for Biomolecular Function Simulation, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe, Hyogo, Japan.
Graduate School of Medical Life Science, Yokohama City University, Tsurumi-ku, Yokohama, Kanagawa, Japan.
Biophys J. 2019 Jan 8;116(1):57-68. doi: 10.1016/j.bpj.2018.11.3122. Epub 2018 Nov 29.
α-amino-3-hydroxy-5-methyl-4-isoaxazolepropionic acid (AMPA) ionotropic glutamate receptors mediate fast excitatory neurotransmission in the central nervous system, and their dysfunction is associated with neurological diseases. Glutamate binding to ligand-binding domains (LBDs) of AMPA receptors induces channel opening in the transmembrane domains of the receptors. The T686A mutation reduces glutamate efficacy so that the glutamate behaves as a partial agonist. The crystal structures of wild-type and mutant LBDs are very similar and cannot account for the observed behavior. To elucidate the molecular mechanism inducing partial agonism of the T686A mutant, we computed the free-energy landscapes governing GluA2 LBD closure using replica-exchange umbrella sampling simulations. A semiclosed state, not observed in crystal structures, appears in the mutant during simulation. In this state, the LBD cleft opens slightly because of breaking of interlobe hydrogen bonds, reducing the efficiency of channel opening. The energy difference between the LBD closed and semiclosed states is small, and transitions between the two states would occur by thermal fluctuations. Evidently, glutamate binding to the T686A mutant induces a population shift from a closed to a semiclosed state, explaining the partial agonism in the AMPA receptor.
α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)离子型谷氨酸受体介导中枢神经系统中的快速兴奋性神经传递,其功能障碍与神经疾病有关。谷氨酸与 AMPA 受体的配体结合域(LBD)结合,诱导受体跨膜域中的通道打开。T686A 突变降低了谷氨酸的效力,使谷氨酸表现为部分激动剂。野生型和突变型 LBD 的晶体结构非常相似,无法解释观察到的行为。为了阐明导致 T686A 突变体部分激动的分子机制,我们使用复制交换伞状采样模拟计算了控制 GluA2 LBD 关闭的自由能景观。在模拟过程中,突变体中出现了一种未在晶体结构中观察到的半封闭状态。在此状态下,由于破坏了叶间氢键,LBD 裂隙略微打开,从而降低了通道打开的效率。LBD 关闭和半封闭状态之间的能量差异很小,通过热波动会发生两种状态之间的转变。显然,谷氨酸与 T686A 突变体结合诱导了从封闭状态到半封闭状态的种群转移,从而解释了 AMPA 受体的部分激动作用。