András Dér

Protein-based integrated optical switching and modulation

The static and dynamic response of optical waveguides coated with a thin protein film of bacteriorhodopsin was investigated. The size and kinetics of the light-induced refractive index changes of the adlayer were determined under different conditions of illumination. The results demonstrate the applicability of this protein as an active, programmable nonlinear optical material in all-optical integrated circuits.

Photoelectric responses in phototactic flagellated algae measured in cell suspension

Abstract A new method for the investigation of electric responses involved in the light reception in microorganisms has been developed. It is based on the detection of photoelectric signals in suspensions of cells (instead of a single cell) by two different techniques: (a) by unilateral excitation of non-oriented cells and (b) after preorientation of the cells (e.g. by gravitaxis or weak, phototactically active light). The method was applied to the flagellated green algae Haematococcus and Chlamydomonas (and several of its mutants). Three main components of the electric signal, which differs in their origin and the mechanisms underlying that registration, were identified. Fast (microsecond) responses reflect charge separation in reaction centres of photosynthesis and are due to the classical light gradient effect on unilateral flash excitation. The later components of the electric signal are involved in photoreception and represent the photoreceptor potential of phototaxis and the calcium-dependent regenerative response. They are measured because of the directional sensitivity of the photoreceptor antenna and the asymmetry of localization of the electric currents involved in the sensory transduction chain. A general similarity between the electric responses in both organisms shows that the sensory transduction chain of photomovements in Chlamydomonas is similar to that described previously for Haematococcus. The advantages of the proposed method are discussed.

Electro-optical measurements on aqueous suspension of purple membrane from Halobacterium halobium.

The permanent dipole moment, polarizability, and the retinal angle of Halobacterium halobium purple membranes were determined at different pH values. All of the parameters have a maximum between pH 5 and 6. There is a reversal in the direction of the permanent dipole moment near pH 5. The value of permanent dipole moment was determined to be 60 D/protein at pH 6.6, and the value obtained for polarizability was 3 X 10(-28) Fm2/membrane fragment. The retinal angle of all-trans retinal was 0.8 degrees smaller than that of the 13-cis conformation.

Electric Signals during the Bacteriorhodopsin Photocycle, Determined over a Wide pH Range

From the electric signals measured after photoexcitation, the electrogenicity 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 calculated 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 Asp96, from the intracellular part of the protein, is not from a well-determined proton donor. The calculated electrogenicities gave good approximations of all of the details of the measured electric signals.

Fluctuations and the Hofmeister Effect

The Hofmeister effect consists in changes of protein solubility triggered by neutral electrolyte cosolutes. Based on the assumption that salts cause stochastic fluctuations of the free energy barrier profiles, a kinetic theory of this phenomenon is proposed. An exponentially correlated noise, of amplitude proportional to the salt concentration, is added to each energy level, and the time-dependence of the mean protein concentration is calculated. It is found that the theory yields the well-known Setschenow equation if the noise correlation time is low in comparison to the aggregation time constant. Experimental data on salting-in agents are well fitted, whereas, in the case of salting-out cosolutes, two independent dichotomic fluctuations are needed to fit the data. This may result from the fact that, in both cases, the low-concentration regime is dominated by salting-in electrostatic contributions, whereas, at high salt concentrations, electron donor/acceptor interactions become important; these have opposite effects. The theory offers a novel way to metricate Hofmeister effects and also leads to thermodynamic quantities, which account for the influence of salts. The formalism may also be applied to describe kinetic phenomena in the presence of cosolutes.

Bacteriorhodopsin as a possible chloride pump

Purple membranes oriented and immobilized in gel show charge transfer at pH 0.55 if the pH is set by HCl. Current appears as laser flash driven transient and also as continuous current by quasi‐continuous illumination. If the pH value 0.55 is set by H2SO4 continuous current is not observed. The results suggest that bacteriorhodopsin may pump chloride ions at low pH.

Protein-based ultrafast photonic switching

Several inorganic and organic materials have been suggested for utilization as nonlinear optical material performing light-controlled active functions in integrated optical circuits, however, none of them is considered to be the optimal solution. Here we present the first demonstration of a subpicosecond photonic switch by an alternative approach, where the active role is performed by a material of biological origin: the chromoprotein bacteriorhodopsin, via its ultrafast BR->K and BR->I transitions. The results may serve as a basis for the future realization of protein-based integrated optical devices that can eventually lead to a conceptual revolution in the development of telecommunications technologies.

Evidence for Loosening of a Protein mechanism

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Fast integrated optical switching by the protein bacteriorhodopsin

State-of-the-art photonic integration technology is ready to provide the passive elements of optical integrated circuits, based either on silicon, glass or plastic materials. The bottleneck is to find the proper nonlinear optical (NLO) materials in waveguide-based integrated optical circuits for light-controlled active functions. Recently, we proposed an approach where the active role is performed by the chromoprotein bacteriorhodopsin as an NLO material, that can be combined with appropriate integrated optical devices. Here we present data supporting the possibility of switching based on a fast photoreaction of bacteriorhodopsin. The results are expected to have important implications for photonic switching technology.

Charge Motion during the Photocycle of Bacteriorhodopsin

The function of bacteriorhodopsin in Halobacterium salinarum is to pump protons from the internal side of the plasma membrane to the external after light excitation, thereby building up electrochemical energy. This energy is transduced into biological energy forms. This review deals with one of the methods elaborated for recording the charge transfer inside the protein. In this method the current produced in oriented purple membrane containing bacteriorhodopsin is measured. It is shown that this method might be applied not only to correlate charge motion with the photocycle reactions but also for general problems like effect of water, electric field, and different ions and buffers for the functioning of proteins.