Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, NY 14214, USA.
Proc Natl Acad Sci U S A. 2013 Mar 19;110(12):E1162-8. doi: 10.1073/pnas.1219777110. Epub 2013 Mar 4.
Familial xerocytosis (HX) in humans is an autosomal disease that causes dehydration of red blood cells resulting in hemolytic anemia which has been traced to two individual mutations in the mechanosensitive ion channel, PIEZO1. Each mutation alters channel kinetics in ways that can explain the clinical presentation. Both mutations slowed inactivation and introduced a pronounced latency for activation. A conservative substitution of lysine for arginine (R2456K) eliminated inactivation and also slowed deactivation, indicating that this mutant's loss of charge is not responsible for HX. Fitting the current vs. pressure data to Boltzmann distributions showed that the half-activation pressure, P1/2, for M2225R was similar to that of WT, whereas mutations at position 2456 were left shifted. The absolute stress sensitivity was calibrated by cotransfection and comparison with MscL, a well-characterized mechanosensitive channel from bacteria that is driven by bilayer tension. The slope sensitivity of WT and mutant human PIEZO1 (hPIEZO1) was similar to that of MscL implying that the in-plane area increased markedly, by ∼6-20 nm(2) during opening. In addition to the behavior of individual channels, groups of hPIEZO1 channels could undergo simultaneous changes in kinetics including a loss of inactivation and a long (∼200 ms), silent latency for activation. These observations suggest that hPIEZO1 exists in spatial domains whose global properties can modify channel gating. The mutations that create HX affect cation fluxes in two ways: slow inactivation increases the cation flux, and the latency decreases it. These data provide a direct link between pathology and mechanosensitive channel dysfunction in nonsensory cells.
人类家族性血红细胞增多症 (HX) 是一种常染色体疾病,导致红细胞脱水,从而引起溶血性贫血。这种疾病可追溯到机械敏感离子通道 PIEZO1 中的两个个体突变。每个突变以可解释临床表现的方式改变通道动力学。两种突变均使失活减慢,并引入明显的激活潜伏期。赖氨酸保守取代精氨酸(R2456K)消除了失活,并且还使去激活减慢,表明该突变体的电荷损失不是导致 HX 的原因。将电流与压力数据拟合到 Boltzmann 分布表明,M2225R 的半激活压力 P1/2 与 WT 相似,而位于 2456 位的突变则向左移动。通过共转染并与细菌中 well-characterized 机械敏感通道 MscL 进行比较来校准绝对应力敏感性,MscL 由双层张力驱动。WT 和突变型人 PIEZO1 (hPIEZO1) 的斜率敏感性与 MscL 相似,这意味着在打开过程中平面内面积显著增加了约 6-20 nm(2)。除了单个通道的行为外,hPIEZO1 通道组还可以同时发生动力学变化,包括失活的丧失和激活的长时间(约 200 ms)沉默潜伏期。这些观察结果表明,hPIEZO1 存在于空间域中,其全局特性可以改变通道门控。导致 HX 的突变以两种方式影响阳离子通量:失活减慢增加阳离子通量,潜伏期缩短则减少阳离子通量。这些数据为非感觉细胞中的病理学和机械敏感通道功能障碍之间提供了直接联系。