膜结合蛋白中的亚基计数
Subunit counting in membrane-bound proteins.
作者信息
Ulbrich Maximilian H, Isacoff Ehud Y
机构信息
Department of Molecular and Cell Biology, University of California, Berkeley Material Science & Physical Bioscience Divisions, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
出版信息
Nat Methods. 2007 Apr;4(4):319-21. doi: 10.1038/nmeth1024. Epub 2007 Mar 18.
Abstract
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.
摘要
在不破坏膜环境的情况下，很难确定膜结合蛋白的亚基数量和化学计量比。在此，我们描述了一种单分子技术，通过观察与感兴趣的蛋白质融合的绿色荧光蛋白（GFP）的漂白步骤来计数活细胞膜中蛋白质的亚基。在用非洲爪蟾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N-甲基-D-天冬氨酸受体（NMDA受体）的组成。
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