Halide ion effects on human Ether-à-go-go related gene potassium channel properties.
Halide ion effects on human Ether-à-go-go related gene potassium channel properties.
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卤化物离子对人类 Ether-à-go-go 相关基因钾通道特性的影响。
DOI：
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发表时间：
2013
影响因子：
1.8
通讯作者：
A. Lagrutta
中科院分区：
文献类型：
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作者：
Haoyu Zeng;Bharath Balasubramanian;Jacob R. Penniman;F. Kinose;J. Salata;A. Lagrutta
人类乙醚-去-去相关基因(HERG)钾通道已被广泛用于对抗筛选潜在药物，作为预测临床QT延长的生物标志物。因此，HERG的高通量分析对于候选药物的早期体外筛选是有价值的，以最大限度地减少由于这种潜在的不利心脏风险而导致的后期药物开发失败。我们开发了一种新的方法，利用氟化钾来提高HERG计数器筛选的吞吐量，该方法使用自动膜片钳系统PatchXpress 7000A。在该方法中，用F(-)对内部氯(-)进行50%的∼取代大大提高了成功率，而不会显著改变HERG通道的生物物理性质或损害数据质量。然而，F(-)或其他卤化物离子对HERG通道性质的影响尚未被详细研究。在这项研究中，我们考察了卤化物离子、F(-)或Br(-)完全取代内溶液中的氯(-)的效果。我们发现：(1)F(-)使HERG通道激活的电压依赖性略微向更正的电压方向移动，而BR(-)使其向更负的电压方向移动；(2)BR(-)使HERG的失活-电压关系和通道完全激活的峰位置向更正的电压方向移动；(3)F(-)使HERG激活变慢，而F(-)和BR(-)使通道关闭更快；(4)F(-)和BR(-)对HERG失活动力学均无影响。总之，与Cl(-)相比，F(-)对HERG通道的激活有微妙的影响，而Br2(-)对HERG通道的某些但不是全部的生物物理性质有明显的影响。
The human Ether-à-go-go related gene (hERG) potassium channel has been widely used to counter screen potential pharmaceuticals as a biomarker to predict clinical QT prolongation. Thus, higher throughput assays of hERG are valuable for early in vitro screening of drug candidates to minimize failure in later-stage drug development due to this potentially adverse cardiac risk. We have developed a novel method utilizing potassium fluoride to improve throughput of hERG counter screening with an automated patch clamp system, PatchXpress 7000A. In that method, ∼50% substitution of internal Cl(-) with F(-) greatly increases success rate without substantially altering the biophysical properties of the hERG channel or compromising data quality. However, effect of F(-) or other halide ions on hERG channel properties has not been studied in detail. In this study, we examined effects of complete replacement of Cl(-) in internal solution with halide ions, F(-), or Br(-). We found that (1) F(-) slightly shifts the voltage dependence of hERG channel activation to more positive voltages, while Br(-) shifts it to more negative voltages; (2) Br(-) shifts to more positive voltages both the inactivation-voltage relationship and the peak position of channel full activation of hERG; (3) F(-) slows hERG activation, while both F(-) and Br(-) make the channel close faster; (4) neither F(-) nor Br(-) have any effect on hERG inactivation kinetics. In conclusion, compared to Cl(-), F(-) has subtle effect on hERG activation, while Br(-) has distinct effects on certain, but not all biophysical properties of hERG channel.
氟化物激活 G 蛋白门控毒蕈碱性心房 K 通道的机制。
DOI：
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发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Yatani,A;Brown,AM
通讯作者：
Brown,AM