Department of Biology, Humboldt University Berlin, Berlin, Germany.
Proteomics. 2011 May;11(10):2124-8. doi: 10.1002/pmic.201000558. Epub 2011 Apr 7.
Membrane proteins are crucial for many essential cellular processes. As membrane proteins function in complexes, methods to detect and to characterize membrane protein-protein interactions are undoubtedly required. Therefore, we developed the "Membrane-Strep-tagged protein interaction experiment" (Membrane-SPINE) that combines the specific purification of a Strep-tagged membrane protein with the reversible fixation of protein complexes by formaldehyde cross-linking. In combination with MS analysis, we suggest Membrane-SPINE as a powerful tool to identify unknown interaction partners of membrane proteins in vivo.
膜蛋白对于许多重要的细胞过程至关重要。由于膜蛋白在复合物中发挥作用,因此无疑需要检测和表征膜蛋白-蛋白相互作用的方法。因此,我们开发了“膜-Strep 标签蛋白相互作用实验”(Membrane-SPINE),该实验将 Strep 标签膜蛋白的特异性纯化与通过甲醛交联可逆固定蛋白复合物相结合。结合 MS 分析,我们建议 Membrane-SPINE 作为一种强大的工具,用于鉴定体内膜蛋白的未知相互作用伙伴。