Trent Tony, Miller Justin J, Blumer Kendall J, Bowman Gregory R
Department of Biochemistry, Biophysics, and Chemical Biology, University of Pennsylvania, Philadelphia, PA 19104-6059, United States.
Department of Biochemistry, Biophysics, and Chemical Biology, University of Pennsylvania, Philadelphia, PA 19104-6059, United States.
J Mol Biol. 2025 Mar 13:169084. doi: 10.1016/j.jmb.2025.169084.
Given the prominence of G protein coupled receptors (GPCRs) as drug targets, targeting their immediate downstream effectors, G proteins, could be valuable as an alternative therapeutic strategy. The discovery that the natural product YM-254890 (YM) can arrest uveal melanoma by specifically inhibiting constitutively active Gq/11 demonstrates the potential of such an approach. However, efforts to find other G protein family-specific inhibitors have had limited success. Better understanding the inhibitory mechanism of YM could facilitate efforts to develop other highly specific G protein inhibitors. We hypothesized that differences between the conformational distributions of various G protein isoforms play important roles in determining whether they are targeted by YM. We addressed this hypothesis by building Markov state models (MSMs) from molecular dynamics simulations of Gα subunits and Gαβγ heterotrimers of three G protein isoforms. We find that in the absence of YM, YM-sensitive Gα subunits have a higher probability of adopting conformations similar to the YM-bound state than YM-insensitive isoforms. There is also strong allosteric coupling between the YM and Gβγ-binding interfaces of Gα. This allostery gives rise to positive cooperativity, wherein the presence of Gβγ enhances preorganization for YM binding. We predict that YM acts as an "allosteric glue" that allosterically stabilizes the complex between Gα and Gβγ despite the minimal contacts between YM and Gβγ.
鉴于G蛋白偶联受体(GPCRs)作为药物靶点的突出地位,将其直接下游效应器G蛋白作为靶向目标,作为一种替代治疗策略可能具有重要价值。天然产物YM-254890(YM)可通过特异性抑制组成型活性Gq/11来阻止葡萄膜黑色素瘤,这一发现证明了这种方法的潜力。然而,寻找其他G蛋白家族特异性抑制剂的努力取得的成功有限。更好地理解YM的抑制机制有助于开发其他高度特异性G蛋白抑制剂。我们假设,各种G蛋白亚型的构象分布差异在决定它们是否被YM靶向方面起着重要作用。我们通过对三种G蛋白亚型的Gα亚基和Gαβγ异源三聚体进行分子动力学模拟构建马尔可夫状态模型(MSMs)来验证这一假设。我们发现,在没有YM的情况下,对YM敏感的Gα亚基比不敏感的亚型更有可能采用与YM结合状态相似的构象。Gα的YM和Gβγ结合界面之间也存在强烈的变构偶联。这种变构作用产生了正协同效应,其中Gβγ的存在增强了YM结合的预组织。我们预测,YM作为一种“变构胶水”,尽管YM与Gβγ之间的接触最少,但它能变构稳定Gα和Gβγ之间的复合物。