Skorinkin Andrei, Nistri Andrea, Giniatullin Rashid
Biophysics Sector, International School for Advanced Studies, via Beirut 4, Trieste 34014, Italy.
J Gen Physiol. 2003 Jul;122(1):33-44. doi: 10.1085/jgp.200308825. Epub 2003 Jun 16.
The mode of action of extracellular protons on ATP-gated P2X2 receptors remains controversial as either enhancement or depression of ATP-mediated currents has been reported. By investigating, at different pH, the electrophysiological effect of ATP on P2X2 receptors and complementing it with receptor modelling, the present study suggests a unified mechanism for both potentiation and inactivation of ATP receptors by protons. Our experiments on patch-clamped PC12 cells showed that, on the same cell, mild acidification potentiated currents induced by low ATP concentrations (<0.1 mM) and attenuated responses to high ATP concentrations (>1 mM) with emergence of current fading and rebound. To clarify the nature of the ATP/H+ interaction, we used the Ding and Sachs's "loop" receptor model which best describes the behavior of such receptors with two open states linked via one inactivated state. No effects by protons could be ascribed to H+-mediated open channel block. However, by assuming that protons facilitated binding of ATP to resting as well as open receptors, the model could closely replicate H+-induced potentiation of currents evoked by low ATP doses plus fading and rebound induced by high ATP doses. The latter phenomenon was due to receptor transition to the inactive state. The present data suggest that the high concentration of protons released with ATP (and catecholamines) from secretory vesicles may allow a dual action of H+ on P2X2 receptors. This condition might also occur on P2X2 receptors of central neurons exposed to low pH during ischemia.
细胞外质子对ATP门控P2X2受体的作用方式仍存在争议,因为有报道称ATP介导的电流既增强又减弱。通过在不同pH值下研究ATP对P2X2受体的电生理效应,并辅以受体建模,本研究提出了质子对ATP受体增强和失活的统一机制。我们对膜片钳PC12细胞的实验表明,在同一细胞上,轻度酸化增强了低ATP浓度(<0.1 mM)诱导的电流,并减弱了对高ATP浓度(>1 mM)的反应,出现电流衰减和反弹。为了阐明ATP/H⁺相互作用的本质,我们使用了丁和萨克斯的“环”受体模型,该模型最能描述此类受体的行为,即两个开放状态通过一个失活状态相连。质子的作用不能归因于H⁺介导的开放通道阻断。然而,通过假设质子促进ATP与静息受体和开放受体的结合,该模型可以紧密复制低ATP剂量引起的H⁺诱导的电流增强以及高ATP剂量引起的衰减和反弹。后一种现象是由于受体转变为失活状态。目前的数据表明,分泌囊泡中与ATP(和儿茶酚胺)一起释放的高浓度质子可能使H⁺对P2X2受体产生双重作用。这种情况也可能发生在缺血期间暴露于低pH值的中枢神经元的P2X2受体上。