Electric signals during the bacteriorhodopsin photocycle, determined over a wide pH range

From the electric signals measured after photoexcitation, the electrogenici ty of the photocycle intermediates of bacteriorhodopsin were determined in a pH range of 4.5-9. Current measurements and absorption kinetic signals at five wavelengths were recorded in the time interval from 300 ns to 0.5 s. To fit the data, the model containing sequential intermediates connected by reversible first-order reactions was used. The electrogenicities were calc ulated from the integral of the current signal, by using the time-dependent concentrations of the intermediates, obtained from the fits. Almost all of the calculated electrogenicities were pH independent, suggesting that the charge motions occur inside the protein. Only the N intermediate exhibited pH-dependent electrogenicity, implying that the protonation of Asp(96), fro m the intracellular part of the protein, is not from a well-determined prot on donor. The calculated electrogenicities gave good approximations of all of the details of the measured electric signals.