Ethylene signaling in plants is decisive for ripening and senescing processes of plants. Thus, fundamental knowledge of the ethylene receptor proteins and specific control of the ethylene signaling cascade will be helpful for sustainable provision of biomass. The objective of the one year SEED FUND project EtMeD was the development of an ethylene measuring device to study ethylene receptor proteins from different plant species. First, a prototype measurement cell has been constructed to show the proof of principle. Finally, a measurement system based on a 24-well-microtiterplate should have been developed to allow parallelization and combined measurements of ethylene and fluorescence-optical methods. Therefore, two core groups participated in this project. The group of Prof. Büchs provided the engineering expertise to design a miniature ethylene measuring cell with a semiconductor gas sensor, While the core group of Prof. Groth focused on the biochemical work of molecular cloning heterologous expression and purification of ethylene receptor proteins.
Despite numerous troubles regarding the long-term stability of the semiconductor gas-sensor and the measurement system, the miniature measurement cell was successfully constructed and the semiconductor gas-sensors could be evaluated and valuable experiences of the behavior of semiconductor gas-sensors could be gained, but measurements with receptor proteins could not be performed in time.
Expression and purification protocols for ethylene receptors were adapted to obtain higher protein amounts. Furthermore, functional folding and structural integrity of the purified recombinant receptor proteins from Arabidopsis and tomato were demonstrated by binding studies with the essential copper metal-cofactor that is required for ethylene binding and biological function of the receptors. Proper storage conditions for long term storage of purified receptors and suitable additives to stabilize the purified detergent solubilized receptors were identified by native differential scanning fluorimetry.
Participating Core Groups
Prof. Dr.-Ing. J. Büchs, AVT - Biochemical Engineering, RWTH Aachen University
Prof. Dr. G. Groth, Biochemical Plant Physiology, HHU Düsseldorf; (Sustainable plant production and resource stewardship)
Contact person / Coordinator
Prof. Dr.-Ing. J. Büchs
RWTH Aachen University
AVT - Biochemical Engineering
Phone: +49 (0)241 / 80 - 24633
Fax: +49 (0)241 / 80 - 22570
01.08.2014 - 31.07.2015
EtMeD is part of the NRW-Strategieprojekt BioSC and thus funded by the Ministry of Innovation, Science and Research of the German State of North Rhine-Westphalia.