Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom; Department of Physiology, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
FEBS Lett. 2014 Jun 5;588(12):2125-8. doi: 10.1016/j.febslet.2014.04.048. Epub 2014 May 8.
Seven P2X purinergic receptor subunits have been identified: P2X1-P2X7. The overlapping expression of P2X2, P2X4 and P2X6 subunits has been shown in different cell types, and functional analysis of P2X receptors in Leydig cells suggests that the three subunits might interact. Here, His6-tagged P2X2, HA-tagged P2X4 and FLAG-tagged P2X6 subunits were co-expressed in tsA 201 cells. After sequential co-immunoprecipitation using anti-HA and anti-FLAG beads, all three subunits were present, demonstrating their interaction. Atomic force microscopy (AFM) imaging revealed receptors that were specifically decorated by both an anti-His6 antibody and an anti-HA Fab fragment, indicating the presence of a P2X2/4/6 heterotrimer. To our knowledge, this is the first report of a P2X receptor containing three different subunits.
已经鉴定出七种 P2X 嘌呤能受体亚基：P2X1-P2X7。在不同的细胞类型中已经显示出 P2X2、P2X4 和 P2X6 亚基的重叠表达，并且对 Leydig 细胞中 P2X 受体的功能分析表明这三个亚基可能相互作用。在这里，His6 标记的 P2X2、HA 标记的 P2X4 和 FLAG 标记的 P2X6 亚基在 tsA 201 细胞中共表达。使用抗 HA 和抗 FLAG 珠粒进行顺序共免疫沉淀后，所有三个亚基都存在，证明它们相互作用。原子力显微镜 (AFM) 成像显示，受体质子化抗体和抗 HA Fab 片段特异性修饰，表明存在 P2X2/4/6 异三聚体。据我们所知，这是第一个包含三个不同亚基的 P2X 受体的报告。
