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SPINE: A method for the rapid detection and analysis of protein–protein interactions in vivo

2007; Wiley; Volume: 7; Issue: 22 Linguagem: Inglês

10.1002/pmic.200700491

1615-9861

Christina Herzberg, Lope Andrés Flórez Weidinger, Bastian Dörrbecker, Sebastian Hübner, Jörg Stülke, Fabian M. Commichau,

Advanced Proteomics Techniques and Applications

Abstract The detection and analysis of protein–protein interactions is one of the central tasks of proteomics in the postgenomic era. For this purpose, we present a procedure, the Strep–protein interaction experiment (SPINE) that combines the advantages of the Strep‐tag protein purification system with those of reversible in vivo protein crosslinking by formaldehyde. Using two Bacillus subtilis regulator proteins, we demonstrate that this method is well suited to isolate protein complexes with high purity and virtually no background. Plasmids allowing the high‐level expression of proteins carrying an N‐ or C‐terminal Strep‐tag in B. subtilis were constructed.