Zhang Mei, Korolkova Yuliya V, Liu Jie, Jiang Min, Grishin Eugene V, Tseng Gea-Ny
Department of Physiology, Virginia Commonwealth University, Richmond 23298, USA.
Biophys J. 2003 May;84(5):3022-36. doi: 10.1016/S0006-3495(03)70028-9.
Peptide toxins with disulfide-stabilized structures have been used as molecular calipers to probe the outer vestibule structure of K channels. We want to apply this approach to the human ether-a-go-go-related gene (HERG) channel, whose outer vestibule is unique in structure and function among voltage-gated K channels. Our focus here is BeKm-1, a HERG-specific peptide toxin that can suppress HERG in the low nM concentration range. Although BeKm-1 shares the three-dimensional scaffold with the well-studied charybdotoxin, the two use different mechanisms in suppressing currents through their target K channels. BeKm-1 binds near, but not inside, the HERG pore, and it is possible that BeKm-1-bound HERG channels can conduct currents although with markedly altered voltage-dependence and kinetics of gating. BeKm-1 and ErgTx1 differ in three-dimensional scaffold, but the two share mechanism of action and have overlapping binding sites on the HERG channel. For both, residues in the middle of the S5-P linker (the putative 583-597 helix) and residues at the pore entrance are critical for binding, although specific contact points vary between the two. Toxin foot printing using BeKm-1 and ErgTx1 will likely provide complementary information about the unique outer vestibule structure of the HERG channel.
具有二硫键稳定结构的肽毒素已被用作分子卡尺来探测钾通道的外部前庭结构。我们希望将这种方法应用于人类醚 - 去极化相关基因(HERG)通道,其外部前庭在电压门控钾通道中具有独特的结构和功能。我们这里关注的是BeKm - 1,一种HERG特异性肽毒素,它能在低纳摩尔浓度范围内抑制HERG。尽管BeKm - 1与经过充分研究的蝎毒素具有相同的三维支架结构,但二者在抑制通过其靶标钾通道的电流时采用不同的机制。BeKm - 1结合在HERG孔道附近而非内部,并且有可能结合了BeKm - 1的HERG通道能够传导电流,尽管其电压依赖性和门控动力学明显改变。BeKm - 1和ErgTx1在三维支架结构上有所不同,但二者作用机制相同,且在HERG通道上具有重叠的结合位点。对于二者而言,S5 - P连接子中间的残基(假定的583 - 597螺旋)和孔道入口处的残基对于结合至关重要,尽管二者的具体接触点有所不同。使用BeKm - 1和ErgTx1进行毒素足迹分析可能会提供有关HERG通道独特外部前庭结构的互补信息。