Carnally Stewart M, Dev Harveer S, Stewart Andrew P, Barrera Nelson P, Van Bemmelen Miguel X, Schild Laurent, Henderson Robert M, Edwardson J Michael
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
Biochem Biophys Res Commun. 2008 Aug 8;372(4):752-5. doi: 10.1016/j.bbrc.2008.05.100. Epub 2008 Jun 2.
There has been confusion about the subunit stoichiometry of the degenerin family of ion channels. Recently, a crystal structure of acid-sensing ion channel (ASIC) 1a revealed that it assembles as a trimer. Here, we used atomic force microscopy (AFM) to image unprocessed ASIC1a bound to mica. We detected a mixture of subunit monomers, dimers and trimers. In some cases, triple-subunit clusters were clearly visible, confirming the trimeric structure of the channel, and indicating that the trimer sometimes disaggregated after adhesion to the mica surface. This AFM-based technique will now enable us to determine the subunit arrangement within heteromeric ASICs.
关于离子通道退化蛋白家族的亚基化学计量学一直存在混淆。最近,酸敏感离子通道(ASIC)1a的晶体结构显示它组装成三聚体。在这里,我们使用原子力显微镜(AFM)对结合在云母上的未处理ASIC1a进行成像。我们检测到亚基单体、二聚体和三聚体的混合物。在某些情况下,三聚体亚基簇清晰可见,证实了通道的三聚体结构,并表明三聚体在粘附到云母表面后有时会解聚。这种基于AFM的技术现在将使我们能够确定异源ASIC内的亚基排列。