Unlocking the Potential of K2P Potassium Channels with Nanobodies

K2P通道对几乎所有人类细胞中发现的基本电流都有贡献。它们受酸、麻醉剂和小分子等多种刺激调节，是治疗疼痛、偏头痛和各种心肺疾病等许多不同疾病的有吸引力的治疗靶点。然而，由于缺乏可在许多活细胞和组织中分析它们的选择性工具，我们充分了解它们的功能特性和作用的能力受到严重阻碍。此外，缺乏选择性的、经过充分验证的针对K2P通道的抗体，这意味着关于它们在全身的表达谱和定位的令人信服的数据很少。研究活组织中任何离子通道的另一个主要复杂性是它们通常由多个亚基组成，这些亚基可以与相关成员聚集在一起形成新的“异质”组合。这种异质多分子化的“混合和匹配”原则对于在活细胞中产生多样性很重要，但往往使分析变得复杂。因此，迫切需要针对这些通道，特别是异质K2P组合的更好和更有选择性的探针。纳米抗体是一种非常小的抗体，来源于骆驼和鲨鱼产生的一种不寻常的抗体。纳米体现在可以很容易地操纵，并在细菌中大量生产。因此，在寻找检测和/或操纵细胞和组织中的K2P通道的新方法时，它们代表了一种令人兴奋的替代小分子化学物质。我们现在有大量的初步数据证明了我们提出的方法的可行性，因此打算使用这些新的探针来解开这些重要离子通道的结构、功能和生理作用。

K2P channels contribute to the basic electrical currents found in nearly all human cells. They are regulated by diverse stimuli, including acid, anaesthetics and small molecules, and represent attractive therapeutic targets for treatment of many different diseases including pain, migraine and various heart & lung disorders.However, our ability to fully understand their functional properties and role is severely hampered by a lack of selective tools that can be used analyse them in many living cells and tissues. Furthermore, a lack of selective, well validated antibodies against K2P channels means there is little convincing data on their expression profiles and localisation throughout the body.Another major complication of studying any ion channel in living tissues is that they often are made up of multiple subunits which can coassemble with related members to form novel 'heteromeric' combinations. This 'mix and match' principle of heteromultimerisation is important for generating diversity in living cells, but often complicates analysis. Better and more selective probes against these channels, especially heteromeric K2P combinations, are therefore desperately needed.Nanobodies are very small antibodies derived from an unusual type of antibody produced by camelids and sharks. Nanobodies can now be easily manipulated and produced in large quantities in bacteria. They therefore represent an exciting alternative to small molecule chemicals when searching for new methods to detect and/or manipulate K2P channels in cells and tissues.We now have extensive preliminary data demonstrating the feasibility of our proposed approach and so intend to use these new probes to unlock the structure, function and physiological role of these important ion channels.