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Technology
SPINE: A method for the rapid detection and analysis of protein–protein interactions in vivo
Article first published online: 12 NOV 2007
DOI: 10.1002/pmic.200700491
Copyright © 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Additional Information
How to Cite
Herzberg, C., Weidinger, L. A. F., Dörrbecker, B., Hübner, S., Stülke, J. and Commichau, F. M. (2007), SPINE: A method for the rapid detection and analysis of protein–protein interactions in vivo. Proteomics, 7: 4032–4035. doi: 10.1002/pmic.200700491
Publication History
- Issue published online: 12 NOV 2007
- Article first published online: 12 NOV 2007
- Manuscript Received: 26 MAY 2007
Funded by
- Deutsche Forschungsgemeinschaft
- Fonds der Chemischen Industrie
- Abstract
- References
- Cited By
Keywords:
- Bacillus subtilis;
- Crosslinking;
- Protein–protein interaction;
- SPINE;
- Strep-tag
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.