Kim Dae In, Jensen Samuel C, Noble Kyle A, Kc Birendra, Roux Kenneth H, Motamedchaboki Khatereh, Roux Kyle J
Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD 57104.
Department of Biological Science, Florida State University, Tallahassee, FL 32306.
Mol Biol Cell. 2016 Apr 15;27(8):1188-96. doi: 10.1091/mbc.E15-12-0844. Epub 2016 Feb 24.
The BioID method uses a promiscuous biotin ligase to detect protein-protein associations as well as proximate proteins in living cells. Here we report improvements to the BioID method centered on BioID2, a substantially smaller promiscuous biotin ligase. BioID2 enables more-selective targeting of fusion proteins, requires less biotin supplementation, and exhibits enhanced labeling of proximate proteins. Thus BioID2 improves the efficiency of screening for protein-protein associations. We also demonstrate that the biotinylation range of BioID2 can be considerably modulated using flexible linkers, thus enabling application-specific adjustment of the biotin-labeling radius.
生物识别（BioID）方法利用一种具有泛素连接活性的生物素连接酶来检测活细胞中的蛋白质-蛋白质相互作用以及邻近蛋白。在此，我们报告了以BioID2为核心对BioID方法的改进，BioID2是一种体积显著更小的具有泛素连接活性的生物素连接酶。BioID2能够更具选择性地靶向融合蛋白，所需的生物素补充量更少，并且对邻近蛋白的标记能力更强。因此，BioID2提高了蛋白质-蛋白质相互作用筛选的效率。我们还证明，使用柔性接头可以显著调节BioID2的生物素化范围，从而能够针对特定应用调整生物素标记半径。
