Photocycle of Dried Acid Purple Form of Bacteriorhodopsin

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Presentation transcript:

Photocycle of Dried Acid Purple Form of Bacteriorhodopsin Géza I. Groma, Lóránd Kelemen, Ágnes Kulcsár, Melinda Lakatos, György Váró Biophysical Journal Volume 81, Issue 6, Pages 3432-3441 (December 2001) DOI: 10.1016/S0006-3495(01)75975-9 Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 1 The spectrum of dried BR and APB. Measuring conditions: 20°C and 51% relative humidity. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 2 FT Raman spectra of dried BR and APB. The insets show the decomposition of the main peak with two Lorentzians, corresponding to all-trans (1526cm−1) and 13-cis (1535–1540cm−1) retinal. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 3 Absorption kinetics measured on dried BR and APB. Measuring conditions were the same as in Fig. 1. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 4 SVD-filtered difference spectra measured on APB at the indicated time after the photoexcitation. Measuring conditions were the same as in Fig. 1. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 5 The spectra of intermediates calculated from the difference spectra shown in Fig. 4. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 6 (A) The fit of the sequential photocycle model (dashed lines) to the time-dependent absorption kinetic signals (continuous lines). (B) Time evolution of the concentration of intermediates. The error of the fit was ±3%. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 7 The free energy, enthalpy, and entropic energy diagram of the APB calculated from the Eyring plots of the temperature-dependent absorption kinetic signals. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 8 Time-dependent absorption kinetic signals measured on APB at different humidities. The arrows show the direction of the decreasing relative humidity: 65, 51, 43, 33, 22, and 12%. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 9 (A) Absorption kinetic signals measured at 610nm in a double excitation experiment: (Y) the signal measured, when only the Nd:YAG laser was fired; (N) the signal measured when only the dye laser was fired; and (Y+N) the signal measured after the double flash, when the dye laser is retarded with 12μs. (B) The resultant signal corresponding to the excitation of the intermediates present at three different time delays after the photoexcitation. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions

Figure 10 Photoelectric signals measured on dried oriented BR and oriented APB. Measuring conditions were the same as in Fig. 1. Biophysical Journal 2001 81, 3432-3441DOI: (10.1016/S0006-3495(01)75975-9) Copyright © 2001 The Biophysical Society Terms and Conditions