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Start of funding 01.07.2007
Potential of high pressure to quantify protein-protein interaction of membrane proteins in vivo
Prof. Dr. Rudi F. Vogel
Technische Universität München
Lehrstuhl für Technische Mikrobiologie
Prof. Dr. Douglas Bartlett
University of California, San Diego
Scripps Institution of Oceanography
The structure and function of membrane proteins is affected by the biophysical state of the bacterial membrane. At the same time, membrane phase and lateral organization of the membrane are influenced by the integrated proteins. This strong interaction of the membrane with its integrated proteins is influenced by pressure and temperature. High hydrostatic pressure (HHP) is a powerful tool to affect the biophysical status of membrane lipids and can therefore be used to study this interaction. In the group of Prof. Vogel, an expression cassette based on ToxR from P. profundum was constructed to study the effect of HHP on membrane protein interaction in vivo. This expression cassette was transformed in a reporter strain of E. coli, which produces ß-galactosidase under the control of ToxR. ToxR activity requires a dimeric state. It was demonstrated in E. coli, that the formation of ToxR dimers can be influenced by HHP as it induces biophysical changes in the membrane structure. In the proposed BaCaTec funded cooperation project it is planned to use a similar expression cassette in P. profundum. The genetic constructions with P. profundum shall be performed most effectively in the Bartlett group because of their long experience with P. profundum.
Final report:
In this project an expression cassette was constructed and transferred to Photobacterium profundum (a deed sea microbe) to determine the effect of high pressure on the expression of the membrane bound signalling protein ToxR. This was achieved during a scientific mission to the Scripps Institution of Oceanography (Prof. Douglas Bartlett) between 15. September and 31. October 2007. This collaboration provided an opportunity for both junior and senior scientists to share and exchange scientific techniques and expertise between laboratories of essentially different disciplines, i.e. food microbiology (high pressure treatment of foods) and deep sea ecology, sharing their interest in high pressure bioscience. In the following a scientist of the Scripps Institution visited TU-München for high pressure microscopy studies. This work formed the basics for a study continued at TU München, which resulted in a number of publications and symposia presentations.
1. Vogel, R. F., Linke, K., Teichert, H., Ehrmann, M. A. 2008. High pressure modulated transport and signalling functions of membrane proteins in models and in vivo. J. Phys.: Conf. Ser. 121 Volume 121 (2008) 112005
2. Vogel, R. F., Ehrmann, M. A. 2008. High pressure sensitive molecular targets in bacteria. Book of Abstracts, VAAM, 9.-11.3.2008, Frankfurt, FGB 03
3. Vogel, R. F., Teichert, H., Linke, K., Periasamy, N., Winter, R., Ehrmann, M. A. 2008. High pressure modulated transport and signaling functions of membrane proteins in model systems and in vivo. Book of Abstracts, VAAM, 9.-11.3.2008, Frankfurt, B0086
4. Linke, K., Periasamy, N., Ehrmann, M. A., Winter, R., and Vogel, R. F. 2008. Influence of pressure on the dimerization of membrane proteins with different transmembrane segments, Appl. Environ. Microbiol. 74, 7821-7823.
5. Vogel, R. F., Linke, K., Teichert, H., Periasamy, N., Ehrmann, M., Winter, R. 2008, High pressure modulated transport and signalling functions of membrane proteins. Conference Proceedings, High Pressure Bioscience and Biotechnology, 14.-17.9.2008, San Diego.
6. Eloe, E. A., Lauro, F. M., Vogel, R. F., and Bartlett, D. H. 2008. The deep-sea bacterium Photobacterium profundum SS9 utilizes separate flagellar systems for swimming and swarming under high pressure conditions. Appl. Environ. Microbiol. 74, 6298-6305.