Crean Rory M, Corbella Marina, Calixto Ana R, Hengge Alvan C, Kamerlin Shina C L
Department of Chemistry - BMC, Uppsala University, Uppsala, Sweden.
Departament de Química Inorgànica i Orgànica (Secció de Química Orgànica) & Institut de Química Teòrica i Computacional (IQTCUB), Universitat de Barcelona, Barcelona, Spain.
QRB Discov. 2024 Jan 24;5:e4. doi: 10.1017/qrd.2024.3. eCollection 2024.
Protein tyrosine phosphatases (PTPs) are crucial regulators of cellular signaling. Their activity is regulated by the motion of a conserved loop, the WPD-loop, from a catalytically inactive open to a catalytically active closed conformation. WPD-loop motion optimally positions a catalytically critical residue into the active site, and is directly linked to the turnover number of these enzymes. Crystal structures of chimeric PTPs constructed by grafting parts of the WPD-loop sequence of PTP1B onto the scaffold of YopH showed WPD-loops in a wide-open conformation never previously observed in either parent enzyme. This wide-open conformation has, however, been observed upon binding of small molecule inhibitors to other PTPs, suggesting the potential of targeting it for drug discovery efforts. Here, we have performed simulations of both enzymes and show that there are negligible energetic differences in the chemical step of catalysis, but significant differences in the dynamical properties of the WPD-loop. Detailed interaction network analysis provides insight into the molecular basis for this population shift to a wide-open conformation. Taken together, our study provides insight into the links between loop dynamics and chemistry in these YopH variants specifically, and how WPD-loop dynamic can be engineered through modification of the internal protein interaction network.
蛋白质酪氨酸磷酸酶(PTPs)是细胞信号传导的关键调节因子。它们的活性受一个保守环(WPD环)从催化无活性的开放构象到催化活性的闭合构象的运动调节。WPD环的运动将一个催化关键残基最佳地定位到活性位点,并与这些酶的周转数直接相关。通过将PTP1B的WPD环序列部分嫁接到YopH支架上构建的嵌合PTPs的晶体结构显示,WPD环处于一种在两种亲本酶中都从未观察到的大开构象。然而,在小分子抑制剂与其他PTPs结合时观察到了这种大开构象,这表明针对它进行药物研发的潜力。在这里,我们对这两种酶都进行了模拟,结果表明催化化学步骤中的能量差异可以忽略不计,但WPD环的动力学性质存在显著差异。详细的相互作用网络分析揭示了这种向大开构象转变的分子基础。总之,我们的研究特别深入了解了这些YopH变体中环动力学与化学之间的联系,以及如何通过修饰内部蛋白质相互作用网络来设计WPD环动力学。