Trapping magnetically oriented chloroplast thylakoid membranes in gels for electric measurements

Electrochromic absorbance change and light gradient photovoltage measurements were carried out in chloroplast thylakoid membranes embedded in different compositions of gels. The goal was to find a system suitable for determining the dependence of the amplitude of the anomalous light gradient photovoltage signal, with opposite sign with respect to the ordinary signal, on the alignment of membranes. We found that polyacrylamide gel drastically increased the permeability of membranes which rendered electric measurements impossible in this gel. Agarose gel was successfully used in electric measurements. We will show that the light gradient photovoltage induced by a 580 nm laser flash depends strongly on the alignment of membranes with respect to the direction of the excitation beam. With face-aligned membranes the amplitude of the anomalous (negative) signal was drastically diminished, and the ordinary (positive) signal became clearly discernible. The observed differences in the decay of the two signals are tentatively explained by differences in the rates of ionic equilibration between different subcompartments of the inner aqueous phase of the thylakoid membrane system.