Subunit counting in membrane-bound proteins
Subunit counting in membrane-bound proteins
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DOI：
10.1038/nmeth1024
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发表时间：
2007-04-01
期刊：
影响因子：
48
通讯作者：
Isacoff, Ehud Y.
中科院分区：
文献类型：
--
作者：
Ulbrich, Maximilian H.;Isacoff, Ehud Y.
膜结合蛋白的亚基数目和化学计量很难在不破坏其膜环境的情况下确定。在这里，我们描述了一种单分子技术，通过观察融合到感兴趣蛋白质的GFP的漂白步骤来计数活细胞膜中的蛋白质亚单位。在用非洲爪哇卵母细胞中表达的已知化学计量比的蛋白质测试该方法后，我们解析了由NR1和NR3亚基组成的NMDA受体的组成。
The subunit number and stoichiometry of membrane-bound proteins are difficult to determine without disrupting their membrane environment. Here we describe a single-molecule technique for counting subunits of proteins in live cell membranes by observing bleaching steps of GFP fused to a protein of interest. After testing the method with proteins of known stoichiometry expressed in Xenopus laevis oocytes, we resolved the composition of NMDA receptors composed of NR1 and NR3 subunits.