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E lectric signals to reveal ion pump function. András Dér. Biological Research Center of the Hungarian Academy of Sciences Institute of Biophysics. P. O. B. 521, H-6701 Szeged, Hungary. Electric signals. Crucial role in life functions:. Signal and energy transduction. Signal transduction.
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Electric signals to revealion pump function András Dér Biological Research Center of the Hungarian Academy of Sciences Institute of Biophysics P. O. B. 521, H-6701 Szeged, Hungary
Electric signals Crucial role in life functions: Signal and energy transduction
Signal transduction Propagation of the nerve impulse Hodgkin, Huxley, Katz Nobel prize (1963)
Energy transduction mitochondrial electron transfer chemiosmosis: P. Mitchell, Nobel prize, 1978 ATP-ase: Boyer, Walker, Skou, Nobel prize, 1997
Why should we measure electric signals? Direct information about kinetics, ion specificity Together with other methods: details of the molecular mechanism is expected to be revealed Physicist's approach: atomic level description - chance to design molecules for biotechnology
Microelectrode techniques fail for most pump proteins How to measure electric signals? Patch clamp; Nobel prize 1991: Neher and Sackmann Alternative methods. Prerequisite: electrically asymmetric sample
1. Surface methods BLM method(Dancsházy et al., 1976; Bamberg et al., 1980) SSM method(Fendler et al., 1992) Advantage: ion specificity Disadvantage: limited spatio-temporal information
2. Bulk methods Suspension method(Keszthelyi and Ormos, 1980) Gel method (Dér et al., 1985) Dried samples(Nagy, 1978; Váró, 1983) Advantageous for kinetic experiments
bR plays a model role among ion-transporting membrane proteins stability, absorption changes, photoelectric effects
cont. light source monochromator (mirror) electrodes transient recorder exciting laser polarizer detector sample trigger amplifier computer input 1 (mirror) input 2 monochromator detector Gel method
Conditions:speed, linearity, insensitivity to geometric details
Properties of ionic relaxation 1. speed 2. anisotropy (F) 3. linearity (E) 4. insensitivity to geometric details (B,C,D)
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Detection of the 3D electric signals Dér et al. (1999)
Testing MD models Measurement Model
Further application examples of the bulk methods Cl- pumping (halorhodopsin, bacteriorhodopsin) Signal transduction (Chlamydomonas rhodopsin, squid rhodopsin) Primary processes of photosynthesis (plant and bacterial photosynthetic reaction centers) Bioelectronics - fast photodiode, motion sensitive camera (bacteriorhodospin)
Acknowledgements Lajos Keszthelyi Pál Ormos György Váró Rudolf Tóth-Boconádi László Oroszi László Fábián Szeged Stefka Taneva Sofia Sándor Suhai Nicoleta Bondar Heidelberg Walther Stoeckenius San Francisco