Single Molecule Biophysics, Institute for Physical and Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue-Strasse 7, 60438 Frankfurt am Main (Germany).
Current address: UCL Medical School, Gower St., London (UK).
Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12049-52. doi: 10.1002/anie.201505664. Epub 2015 Aug 20.
Techniques based on fluorescence microscopy are increasingly used to count proteins in cells, but few stoichiometrically well-defined standards are available to test their accuracy. A selection of bacterial homo-oligomers were developed that contain 10-24 subunits and fully assemble when expressed in mammalian cells, and they can be used to easily validate/calibrate molecular counting methods. The utility of these standards was demonstrated by showing that nuclear pores contain 32 copies of the Nup107 complex.
基于荧光显微镜的技术越来越多地被用于细胞内蛋白质的计数,但很少有化学计量定义明确的标准可用于测试其准确性。我们开发了一系列包含 10-24 个亚基的细菌同聚寡聚体,这些寡聚体在哺乳动物细胞中表达时会完全组装,可用于轻松验证/校准分子计数方法。通过显示核孔含有 32 个核孔蛋白 107 复合物拷贝,证明了这些标准的实用性。