Barrera Nelson P, Shaifta Yasin, McFadzean Ian, Ward Jeremy P T, Henderson Robert M, Edwardson J Michael
Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
Biochem Biophys Res Commun. 2007 Jul 13;358(4):1086-90. doi: 10.1016/j.bbrc.2007.05.039. Epub 2007 May 15.
We have determined the subunit stoichiometry of the transient receptor potential C1 (TRPC1) channel by imaging isolated channels using atomic force microscopy (AFM). A frequency distribution of the molecular volumes of individual channel particles had two peaks, at 170 and 720 nm(3), corresponding with the expected sizes of TRPC1 monomers and tetramers, respectively. Complexes were formed between TRPC1 channels and antibodies against a V5 epitope tag present on each subunit. The frequency distribution of angles between pairs of bound antibodies had two peaks, at 88 degrees and 178 degrees. This result again indicates that the channel assembles as a tetramer.
我们通过使用原子力显微镜（AFM）对分离出的通道进行成像，确定了瞬时受体电位C1（TRPC1）通道的亚基化学计量。单个通道颗粒的分子体积频率分布有两个峰值，分别在170和720 nm³处，分别对应于TRPC1单体和四聚体的预期大小。TRPC1通道与针对每个亚基上存在的V5表位标签的抗体之间形成了复合物。结合抗体对之间角度的频率分布有两个峰值，分别在88度和178度。这一结果再次表明该通道组装为四聚体。
