WorldWideScience

Sample records for bacteriorhodopsins

  1. Absorption characteristics of bacteriorhodopsin molecules

    Indian Academy of Sciences (India)

    H K T Kumar; K Appaji Gowda

    2000-03-01

    The bacteriorhodopsin molecule absorbs light and undergoes a series of structural transformation following a well-defined photocycle. The complex photocycle is transformed to an equivalent level diagram by considering the lifetime of the intermediate states. Assuming that only and states are appreciably populated at any instant of time, the level diagram is further simplified to two-level system. Based on the rate equations for two-level system, an analytic expression for the absorption coefficient of bacteriorhodopsin molecule is derived. It is applied to study the behaviour of absorption coefficient of bacteriorhodopsin film in the visible wavelength region of 514 nm. The dependence of absorption coefficient of bacteriorhodopsin film on the thickness of the film, total number density of active molecules and initial number density of molecules in -state is presented in the graphical form.

  2. Nonlinear optical properties of bacteriorhodopsin

    Science.gov (United States)

    Hendrickx, Eric; Verbiest, Thierry; Clays, Koen J.; Persoons, Andre P.

    1993-04-01

    In this paper we show the applicability of Hyper-Rayleigh scattering to obtain hyperpolarizabilities of ionic and biochemical compounds. It was found that dark-adapted bacteriorhodopsin and its isolated chromophore have considerable second order nonlinear optical properties. Information obtained from depolarization studies of the scattered light is discussed.

  3. Resonant optical rectification in bacteriorhodopsin.

    Science.gov (United States)

    Groma, Géza I; Colonna, Anne; Lambry, Jean-Christophe; Petrich, Jacob W; Váró, György; Joffre, Manuel; Vos, Marten H; Martin, Jean-Louis

    2004-05-25

    The relative role of retinal isomerization and microscopic polarization in the phototransduction process of bacteriorhodopsin is still an open question. It is known that both processes occur on an ultrafast time scale. The retinal trans-->cis photoisomerization takes place on the time scale of a few hundred femtoseconds. On the other hand, it has been proposed that the primary light-induced event is a sudden polarization of the retinal environment, although there is no direct experimental evidence for femtosecond charge displacements, because photovoltaic techniques cannot be used to detect charge movements faster than picoseconds. Making use of the known high second-order susceptibility chi(2) of retinal in proteins, we have used a nonlinear technique, interferometric detection of coherent infrared emission, to study macroscopically oriented bacteriorhodopsin-containing purple membranes. We report and characterize impulsive macroscopic polarization of these films by optical rectification of an 11-fs visible light pulse in resonance with the optical transition. This finding provides direct evidence for charge separation as a precursor event for subsequent functional processes. A simple two-level model incorporating the resonant second-order optical properties of retinal, which are known to be a requirement for functioning of bacteriorhodopsin, is used to describe the observations. In addition to the electronic response, long-lived infrared emission at specific frequencies was observed, reflecting charge movements associated with vibrational motions. The simultaneous and phase-sensitive observation of both the electronic and vibrational signals opens the way to study the transduction of the initial polarization into structural dynamics. PMID:15148391

  4. Nonlinear Optical Studies of Bacteriorhodopsin

    Science.gov (United States)

    Rao, D. V. G. L. N.; Aranda, F. J.; Chen, Z.; Akkara, J. A.; Kaplan, D. L.; Nakashima, M.

    We report interesting results on nonlinear optics at low powers in bacteriorhodopsin films with applications in all-optical switching and modulation. Chemically stabilized films of bacteriorhodopsin in a polymer matrix for which the lifetime of the excited M state is 3 to 4 orders of magnitude longer than that of water solutions of wild-type bR were used in these experiments. Due to the sensitivity of the films, very small powers of order microwatts are required for optical phase conjugation. The influence of the fast photochemical M to B transition induced by blue light on the saturation intensity, phase conjugate intensity and switching time was established. We also report our measurements of the intensity dependence of the self-focusing and self-defocusing properties of wild-type bR in water solution using the Z-scan technique with low power cw lasers at two wavelengths on either side of the absorption band. Our measurements indicate that the sign of the nonlinearity depends on the wavelength and the magnitude depends on the fluence of the incident laser beam. The observed self-focusing and defocusing is not due to the intrinsic electronic nonlinearity. The observations can be explained in terms of the Kramers-Kronig dispersion relation that relates the real and imaginary parts of the complex index of refraction.

  5. Optical limiting by chemically enhanced bacteriorhodopsin films

    Science.gov (United States)

    Song, Q. Wang; Zhang, Chungping; Gross, Richard; Birge, Robert

    1993-05-01

    Measurements of effective nonlinearity of a chemically enhanced bacteriorhodopsin film are presented, using 2-scan method. Optical limiting properties and the film's nonlinear transmission properties of the film are also studied.

  6. Nonlinear Optical Image Processing with Bacteriorhodopsin Films

    Science.gov (United States)

    Downie, John D.; Deiss, Ron (Technical Monitor)

    1994-01-01

    The transmission properties of some bacteriorhodopsin film spatial light modulators are uniquely suited to allow nonlinear optical image processing operations to be applied to images with multiplicative noise characteristics. A logarithmic amplitude transmission feature of the film permits the conversion of multiplicative noise to additive noise, which may then be linearly filtered out in the Fourier plane of the transformed image. The bacteriorhodopsin film displays the logarithmic amplitude response for write beam intensities spanning a dynamic range greater than 2.0 orders of magnitude. We present experimental results demonstrating the principle and capability for several different image and noise situations, including deterministic noise and speckle. Using the bacteriorhodopsin film, we successfully filter out image noise from the transformed image that cannot be removed from the original image.

  7. Novelty filter that uses a bacteriorhodopsin film

    Science.gov (United States)

    Okamoto, Takayuki; Yamaguchi, Ichirou; Boothroyd, Simon A.; Chrostowski, Jacek

    1997-01-01

    We propose a new novelty optical filter that uses a bacteriorhodopsin film. This filter is based on the time-dependent nonlinear diffraction efficiency of real-time holograms recorded in the film. As soon as the signal beam carrying a pattern is diffracted by the polarization hologram recorded in the bacteriorhodopsin film, it begins to erase the hologram and suppresses the diffraction of the beam at the position of the stationary part of the pattern. This filter enhances only leading edges of moving patterns. In this system undesired scattered light, which is orthogonally polarized to the diffracted beam, is discriminated by a polarizer.

  8. Nonlinear transmittance of the 4-keto bacteriorhodopsin

    Science.gov (United States)

    Vanhanen, J.; Leppanen, V. P.; Jaaskelainen, T.; Parkkinen, J. P. S.; Parkkinen, S.

    1999-09-01

    The photocycle of the 4-keto bacteriorhodopsin is investigated. We constructed a multilevel theoretical model for the nonlinear transmittance properties of the material. Adjusting the relaxation parameters we are able to fit the theoretical intensity dependent transmittance curves into the experiments and to determine the photocycle from simple optical measurements.

  9. Nonlinear transmission properties in bacteriorhodopsin-embedded photonic crystal

    Science.gov (United States)

    Okada-Shudo, Yoshiko; Ishihara, Teruya

    2003-11-01

    Transmission spectra and photoinduced transmission change are observed in periodic waveguide which consist of a quartz grating substrate and a thin protein film of bacteriorhodopsin. We propose a scheme to achieve all optical switching using the photoinduced refractive index change of bacteriorhodopsin.

  10. Retinal isomerization dynamics in dry bacteriorhodopsin films

    Science.gov (United States)

    Colonna, Anne; Groma, Géza I.; Vos, Marten H.

    2005-10-01

    The primary photoprocesses in neutral and acid forms of oriented dried bacteriorhodopsin films were investigated by femtosecond absorption spectroscopy. The excitation energy dependence of the signals was used to distinguish photochemistry from processes involving photophysics of photocycle intermediates. Both the kinetics and the quantum yield of all- trans excited state decay by retinal photoisomerization and subsequent J → K transition were found to be very similar as in hydrated environments. Therefore, unlike slower photocycle phases, communication of the retinal with the environment does not play a role in retinal isomerization. Our results are important for understanding recent nonlinear optical applications of such films.

  11. Ultrafast all-optical switching in bacteriorhodopsin

    Science.gov (United States)

    Roy, Sukhdev; Singh, C. P.; Reddy, K. P. J.

    2001-04-01

    All-optical switching has been demonstrated in bacteriorhodopsin based on excited-state nonlinear absorption. A probe laser beam at 640 nm corresponding to the O-state absorption maximum is switched due to a strong pulsed pump laser beam at 570 nm, that corresponds to the maximum ground state absorption. We have studied the effect of variation in pulse width and in small signal absorption coefficient on the switching characteristics. The switching time decreases as the pulse width of the pump beam decreases and the small signal absorption coefficient increases. The switching contrast depends mainly on the peak pumping intensity.

  12. Multiplexed Holographic Data Storage in Bacteriorhodopsin

    Science.gov (United States)

    Mehrl, David J.; Krile, Thomas F.

    1999-01-01

    Biochrome photosensitive films in particular Bacteriorhodopsin exhibit features which make these materials an attractive recording medium for optical data storage and processing. Bacteriorhodopsin films find numerous applications in a wide range of optical data processing applications; however the short-term memory characteristics of BR limits their applications for holographic data storage. The life-time of the BR can be extended using cryogenic temperatures [1], although this method makes the system overly complicated and unstable. Longer life-times can be provided in one modification of BR - the "blue" membrane BR [2], however currently available films are characterized by both low diffraction efficiency and difficulties in providing photoreversible recording. In addition, as a dynamic recording material, the BR requires different wavelengths for recording and reconstructing of optical data in order to prevent the information erasure during its readout. This fact also put constraints on a BR-based Optical Memory, due to information loss in holographic memory systems employing the two-lambda technique for reading-writing thick multiplexed holograms.

  13. Multiplexed Holographic Optical Data Storage In Thick Bacteriorhodopsin Films

    Science.gov (United States)

    Downie, John D.; Timucin, Dogan A.; Gary, Charles K.; Ozcan, Meric; Smithey, Daniel T.; Crew, Marshall

    1998-01-01

    The optical data storage capacity of photochromic bacteriorhodopsin films is investigated by means of theoretical calculations, numerical simulations, and experimental measurements on sequential recording of angularly multiplexed diffraction gratings inside a thick D85N BR film.

  14. Electron microscopic observation and rotational diffusion measurement of bacteriorhodopsin in lipid vesicles

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The morphology of bacteriorhodopsin reconstituted into dimyristoylphosphatidylcholine and egg-phosphatidylcholine vesicles was observed by freeze-fracture electron microscopy. The rotational diffusion of bacteriorhodopsin at different concentrations of melittin was measured by observing flash-induced transient dichroism in dimyristoylphosphatidylcholine vesicles. In the presence of melittin, bacteriorhodopsin molecules in dimyristoylphosphatidylcholine vesicles were aggregated into large particles or patches, and the ability of rotational diffusion of bacteriorhodop sin in vesicles was decreased. This suggests that melittin produces its effect via direct electrostatic interaction with bacteriorhodopsin. Low temperature-induced aggregation of bacteriorhodopsin was also observed in dimyristoylphosphatidylcholine vesicles. Low temperature may cause phase separation. Bacteriorhodopsin was also successfully reconstituted into egg-phosphatidylcholine vesicles, but Iow temperature-induced aggregation of bacteriorhodopsin in dimyristoylphosphati dylcholine cannot appear in egg-phosphatidylcholine vesicles. This suggests that different lipids have different effects on bacteriorhodopsin in vesicles.

  15. Bacteriorhodopsin: Tunable Optical Nonlinear Magnetic Response

    CERN Document Server

    Bovino, F A; Sibilia, C; Giardina, M; Váró, G; Gergely, C

    2011-01-01

    We report on a strong and tunable magnetic optical nonlinear response of Bacteriorhodopsin (BR) under "off resonance" femtosecond (fs) pulse excitation, by detecting the polarization map of the noncollinear second harmonic signal of an oriented BR film, as a function of the input beam power. BR is a light-driven proton pump with a unique photochemistry initiated by the all trans retinal chromophore embedded in the protein. An elegant application of this photonic molecular machine has been recently found in the new area of optogenetics, where genetic expression of BR in brain cells conferred a light responsivity to the cells enabling thus specific stimulation of neurons. The observed strong tunable magnetic nonlinear response of BR might trigger promising applications in the emerging area of pairing optogenetics and functional magnetic resonance imaging susceptible to provide an unprecedented complete functional mapping of neural circuits.

  16. Bacteriorhodopsin-based photochromic pigments for optical security applications

    Science.gov (United States)

    Hampp, Norbert A.; Fischer, Thorsten; Neebe, Martin

    2002-04-01

    Bacteriorhodopsin is a two-dimensional crystalline photochromic protein which is astonishingly stable towards chemical and thermal degradation. This is one of the reasons why this is one of the very few proteins which may be used as a biological pigment in printing inks. Variants of the naturally occurring bacteriorhodopsin have been developed which show a distinguished color change even with low light intensities and without the requirement of UV-light. Several pigments with different color changes are available right now. In addition to this visual detectable feature, the photochromism, the proteins amino acid sequence can be genetically altered in order to code and identify specific production lots. For advanced applications the data storage capability of bacteriorhodopsin will be useful. Write-once-read-many (WORM) recording of digital data is accomplished by laser excitation of printed bacteriorhodopsin inks. A density of 1 MBit per square inch is currently achieved. Several application examples for this biological molecule are described where low and high level features are used in combination. Bacteriorhodopsin-based inks are a new class of optical security pigments.

  17. Numerical simulation of the nonlinear optical response of bacteriorhodopsin

    Science.gov (United States)

    Kowalski, Gregory J.

    1996-10-01

    The numerical simulation of the nonlinear optical behavior of bacteriorhodopsin in a solution of water is described. Relationships for the intensity dependent absorption coefficient and index of refraction are developed and used in the numerical simulation of bacteriorhodopsin as an optical limiter and as defocussing element for laser pulses in the picosecond regime. The algorithm is a transient finite volume method that is coupled with a 'ray model' of the radiation which simultaneously solves the heat transfer and Maxwell's equations. The nonlinear behavior of the material is included in this analysis using a modified Euler predictor-corrector integration technique. Calculated power limiting and z-scan curves are in qualitative agreement with experiments. These results indicate that the code can be used to investigate and optimize optical systems which use the nonlinear behavior of bacteriorhodopsin.

  18. Bacteriorhodopsin overview of fundamentals and applications

    Science.gov (United States)

    Thai, Serey

    1999-07-01

    Bacteriorhodopsin (BR) is a light transducing photochromic protein in the purple membrane of a salt-loving microorganism that inhabits salt marshes. It has strong absorption in a broad region of the visible spectrum. The B- state in the photocycle can be considered to be the ground state, which has absorption maxima at 570 nm. Perhaps, the most intriguing features of this organic photopolymer are its extraordinary stability in the chemical, thermal and photochemical sense, its large optical nonlinearity, dynamic nature, durability, real-time holographic recording capabilities, and information storage potential. Furthermore, BR-doped polymer film can be fabricated for a large-scale application, whereas photorefractive crystals like BSO or KNSBM cannot be grown easily to the same dimension as BR. Hence, BR's potential in optical system includes transient dynamic applications of an M-type hologram and 3D optical memories of a branched photocycle that shows a great promise for data storage and retrieval due to its high capacity. The major advantages of this organic photopolymer include high density, low cost, low weight and portability which are a projected requirement for the Air Force and commercial applications.

  19. Enhancement of photoanisotropy in Bacteriorhodopsin films

    Science.gov (United States)

    Rao, D. V. G. L. N.; Wu, Pengfei

    2003-03-01

    The biological material of Bacteriorhodopsin (bR) and its derivatives are among the most promising candidates for potential applications in photonics in view of their large optical nonlinearity, ease of optimization and tailoring optical properties. We report a novel scheme for significant enhancement of photo-anisotropic effects in bR films using two exciting beams at different wavelengths with orthogonal polarization. We monitor the photoinduced anisotropy with a probe beam passing through the bR film placed between two crossed polarizers. Near twenty times enhancement of probe beam intensity has been observed as compared with the case of only one exciting beam. The mechanism of the enhancement originates from optimization of direction-selected photo-isomerization of the biomaterial controlled by the polarized exciting beams. We also demonstrate an all-optical switch with the additional novel feature of output sign-control by applying this technique. It is possible to achieve fast optical switching since the photo-isomerization of M to B state of the bR molecule may be as fast as nanoseconds.

  20. Measurements of photoinduced refractive index changes in bacteriorhodopsin films

    Indian Academy of Sciences (India)

    Ravinder Kumar Banyal; B Raghavendra Prasad

    2007-03-01

    We report the pump-probe measurements of nonlinear refractive index changes in photochromic bacteriorhodopsin films. The photoinduced absorption is caused by pump beam at 532 nm and the accompanying refractive index changes are studied using a probe beam at 633 nm. The proposed technique is based on a convenient and accurate determination of optical path difference using digital interferometry-based local fringe shift. The results are presented for the wild-type as well as genetically modified D96N variant of the bacteriorhodopsin.

  1. Measurements of photoinduced refractive index changes in bacteriorhodopsin films

    Science.gov (United States)

    Banyal, Ravinder Kumar; Raghavendra Prasad, B.

    2007-03-01

    We report the pump--probe measurements of nonlinear refractive index changes in photochromic bacteriorhodopsin films. The photoinduced absorption is caused by pump beam at 532 nm and the accompanying refractive index changes are studied using a probe beam at 633 nm. The proposed technique is based on a convenient and accurate determination of optical path difference using digital interferometry-based local fringe shift. The results are presented for the wild-type as well as genetically modified D96N variant of the bacteriorhodopsin.

  2. Fast integrated optical switching by the protein bacteriorhodopsin

    Science.gov (United States)

    Fábián, László; Wolff, Elmar K.; Oroszi, László; Ormos, Pál; Dér, András

    2010-07-01

    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.

  3. Optical CDMA system using bacteriorhodopsin for optical data storage

    Science.gov (United States)

    Bae; Yang; Jin; Lee; Park

    1999-11-01

    An optical CDMA (code division multiple access) system for the optical data storage using bacteriorhodopsin (BR) is reported as an application of the BR materials. The desired signal of multiple input can be recorded and reconstructed by use of orthogonal codes. An experimental setup is proposed and demonstrated. PMID:10585180

  4. Optical Processing of Speckle Images with Bacteriorhodopsin for Pattern Recognition

    Science.gov (United States)

    Downie, John D.; Tucker, Deanne (Technical Monitor)

    1994-01-01

    Logarithmic processing of images with multiplicative noise characteristics can be utilized to transform the image into one with an additive noise distribution. This simplifies subsequent image processing steps for applications such as image restoration or correlation for pattern recognition. One particularly common form of multiplicative noise is speckle, for which the logarithmic operation not only produces additive noise, but also makes it of constant variance (signal-independent). We examine the optical transmission properties of some bacteriorhodopsin films here and find them well suited to implement such a pointwise logarithmic transformation optically in a parallel fashion. We present experimental results of the optical conversion of speckle images into transformed images with additive, signal-independent noise statistics using the real-time photochromic properties of bacteriorhodopsin. We provide an example of improved correlation performance in terms of correlation peak signal-to-noise for such a transformed speckle image.

  5. Studies on light transduction by bacteriorhodopsin and rhodopsin

    International Nuclear Information System (INIS)

    The visual photoreceptor pigments in vertebrates and invertebrates all use retinal (vitamin A aldehyde) as the light-absorbing molecule. Recently, Stoeckenius et al. discovered bacteriorhodopsin (bR) in the purple membrane of the extreme halophile, Halobacterium halobium, which also contains all-trans retinal as the chromophore, bR carries out light-dependent proton translocation from the inside to the outside of the H. halobium cell. Since the discovery of bR, H. halobium has been found to elaborate three more retinal-based light-transducing proteins. These are halorhodopsin, a chloride ion pump, and sensory rhodopsins I and II. The authors are carrying out structure-function studies of bacteriorhodopsin, bovine rhodopsin, and related proteins primarily by the technique of recombinant DNA; they summarize below the results they have obtained recently

  6. Nonlinear optical properties of bacteriorhodopsin, retinal, and related molecules

    Science.gov (United States)

    Hendrickx, Eric; Clays, Koen J.; Vinckier, A.; Persoons, Andre P.; Dehu, Christophe; Bredas, Jean-Luc

    1995-10-01

    The first hyperpolarizabilities, (beta) , of bacteriorhodopsin, retinal, and related molecules were determined experimentally by using the hyper-Rayleigh scattering technique and compared to the calculated values obtained with the semiempirical intermediate neglect of differential overlap/configuration interaction/sum-over-states method. The experimental and theoretical results are in excellent mutual agreeement. The hyper-Rayleigh scattering technique is shown to be very sensitive to the degree of solubilization of bacteriorhodopsin. Theoretical and experimental data confirm the expected dependence of (beta) on the first transition energy as well as an exponential increase of (beta) with the number of double bonds. It was found that, upon trans to 13-cis or 9-cis isomerization of a retinal double bond, a constant fraction of the (beta) value is lost, regardless of the nature of the electron withdrawing group or the solvent of choice.

  7. Do Cation-π Interactions Exist in Bacteriorhodopsin

    Institute of Scientific and Technical Information of China (English)

    HU Kun-Sheng; WANG Guang-Yu; HE Jin-An

    2001-01-01

    Metal ions are essential to the structure and physiological functions of bacteriorhodopsin. Experimental evidence suggests the existence of specific cation binding to the negatively charged groups of Asp85 and Asp212 via an electrostatic interaction. However, only using electrostatic force is not enough to explain the role of the metal cations because the carboxylate of Asp85 is well known to be protonated in the M intermediate. Considering the presence of some aromatic amino acid residues in the vicinity of the retinal pocket, the existence of cation-π interactions between the metal cation and aromatic amino acid residues is suggested. Obviously, introduction of this kind of interaction is conducive to understanding the effects of the metal cations and aromatic amino acid residues inside the protein on the structural stability and proton pumping of bacteriorhodopsin.

  8. Bacteriorhodopsin as a Possible Element of Membrane Bioreactors

    Science.gov (United States)

    Maksimychev, A. V.; Chamorovskii, S. K.

    1988-06-01

    Certain approaches to the creation of membrane bioreactors, representing an example of integrated membrane systems, are examined. The characteristic features of the use of organised molecular assemblies in such systems as sensor and regulatory elements are discussed. The properties of the retinal-protein complex of bacteriorhodopsin as a promising component of integrated membrane systems, capable of carrying out regulatory functions, are examined. The bibliography includes 139 references.

  9. Nonlinear polarization interaction in bacteriorhodopsin films with anisotropically saturating absorption

    Science.gov (United States)

    Korchemskaya, Elena Y.; Soskin, Marat S.; Stepanchikov, Dmitriy A.; Druzhko, Anna B.; Dyukova, Tatyana V.

    1996-06-01

    The effect of protein and matrix modifications on the photoanisotropic properties is studied for developing the concept of impact upon the main optical properties of the dynamic optical material based on bacteriorhodopsin (BR) both interaction of transmembrane protein--chromophore complex BR with matrix and interaction of protein opsin with chromophore retinal. Also possibility of the application of BR-films for the light polarization modulator is proposed.

  10. Nonlinear polarization-modulated spectroscopy of bacteriorhodopsin and its analogues

    Science.gov (United States)

    Taranenko, V. B.; Bazhenov, V. Yu; Kulikovskaya, O. A.

    1996-09-01

    We report on a novel nonlinear polarization-modulated spectroscopic method for an accurate measurement of the nonlinear change of both real and imaginary parts of the complex refractive index in isotropic materials having either scalar or tensor photoresponse. It is based on a vector two-wave-mixing interaction and heterodyne detection of dynamic change of optical polarization. New data on steady-state and transient nonlinear characteristics of bacteriorhodopsin-based materials (suspensions and polymer films) are obtained using this method.

  11. All-optical logic-gates based on bacteriorhodopsin film

    Institute of Scientific and Technical Information of China (English)

    Chen Gui-Ying; Zhang Chun-Ping; Guo Zong-Xia; Tian Jian-Guo; Zhang Guang-Yin; Song Qi-Wang

    2005-01-01

    Based on self-diffraction in bacteriorhodopsin (bR) film, we propose all-optical NOT, XOR, half adder and XNOR logic operations. Using the relation between diffraction light and the polarization states of recording beams, we demonstrate NOT and XNOR logic operations. Studying the relation of polarization states among the diffracting, recording and reading beams, we implement XOR logic and half adder operations with three inputs. The methods are simple and practicable.

  12. Three electronic state model of the primary phototransformation of bacteriorhodopsin.

    OpenAIRE

    Humphrey, W; H. Lu; Logunov, I; Werner, H.J.; Schulten, K

    1998-01-01

    The primary all-trans --> 13-cis photoisomerization of retinal in bacteriorhodopsin has been investigated by means of quantum chemical and combined classical/quantum mechanical simulations employing the density matrix evolution method. Ab initio calculations on an analog of a protonated Schiff base of retinal in vacuo reveal two excited states S1 and S2, the potential surfaces of which intersect along the reaction coordinate through an avoided crossing, and then exhibit a second, weakly avoid...

  13. High production of bacteriorhodopsin from wild type Halobacterium salinarum.

    Science.gov (United States)

    Seyedkarimi, Mansooreh-Sadat; Aramvash, Asieh; Ramezani, Rohollah

    2015-09-01

    Bacteriorhodopsin (bR) is a trans-membrane proton pump found in the purple membrane of Halobacterium salinarum. This protein has high photochemical and photoelectric conversion efficiency and thermal stability, allowing it to withstand high temperatures, high salinity, and nutritionally-limited environments. The ability of this protein to convert light energy into chemical energy has applications that are mainly therapeutic/diagnostic and research-oriented. There is increasing demand for bacteriorhodopsin production in different fields. The present study maximized bacteriorhodopsin production using H. salinarum. The physical parameters of illumination, agitation speed, temperature, and nitrogen source were studied using a fractional factorial design to determine the optimal levels of each. The most suitable nitrogen source was determined to be peptone from meat. The optimal temperature was 39 °C, agitation speed was 150 rpm, and light intensity was 6300 lux for bR production. Under these conditions, the maximum bR yield was 196 mg/l, which is about 4.23 fold greater than those obtained with basal medium. The proposed strategies could be used for bR production using this archaeobacterium; the results are the highest reported thus far from a batch culture of H. salinarum. PMID:26254806

  14. Fractal morphological analysis of Bacteriorhodopsin (bR) layers deposited onto Indium Tin Oxide (ITO) electrodes

    International Nuclear Information System (INIS)

    Uniform Bacteriorhodopsin layers for the purpose of fabricating Bacteriorhodopsin-based biosensors were prepared by allowing drying of the layers under a constant electric field. To properly observe and understand the 'electric field effect' on the protein Bacteriorhodopsin, the electric and non-electric field influenced Bacteriorhodopsin layers prepared using a manual syringe-deposition method applied onto Indium Tin Oxide electrodes were structurally investigated using Scanning Electron Microscopy and Atomic Force Microscopy. The results yield obvious morphological differences between the electric and non-electric field assisted Bacteriorhodopsin layers and brings to attention the occurrence of the so-called 'coffee-ring' effect in the latter case. We applied stochastic fractal method based on the generalized Cauchy process to describe the morphological features surrounding the void. Fractal dimension is used to characterize the local regularity of the Bacteriorhodopsin clusters and the correlation exponent is used to describe the long-range correlation between the clusters. It is found that the Bacteriorhodopsin protein tends to exhibit with strong spatial correlation in the presence of external electric field compared to in absence of the electric field. Long-range correlation in the morphological feature may be associated to the enhancement of aggregation process of Bacteriorhodopsin protein in the presence of electric field, thereby inhibiting the formation of the so-called 'coffee-ring' effect. As such, the observations discussed in this work suggest some amount of control of surface uniformity when forming layers.

  15. Rate Constant Change of Photo Reaction of Bacteriorhodopsin Observed in Trimeric Molecular System.

    Science.gov (United States)

    Tsujiuchi, Yutaka; Masumoto, Hiroshi; Goto, Takashi

    2016-04-01

    To elucidate the time evolution of photo reaction of bacteriorhodopsin in glycerol mixed purple membrane at around 196 K under irradiation by red light, a kinetic model was constructed. The change of absorption with irradiation at times of 560 nm and 412 nm was analyzed for the purpose of determining reaction rates of photo reaction of bacteriorhodopsin and its product M intermediate. In this study it is shown that reaction rates of conversion from bacteriorhodopsin to the M intermediate can be explained by a set of linear differential equations. This model analysis concludes that bacteriorhodopsin in which constitutes a trimer unit with other two bacteriorhodopsin molecules changes into M intermediates in the 1.73 of reaction rate, in the initial step, and according to the number of M intermediate in a trimer unit, from three to one, the reaction rate of bacteriorhodopsin into M intermediates smaller as 1.73, 0.80, 0.19 which caused by influence of inter-molecular interaction between bacteriorhodopsin. PMID:27451646

  16. The behaviours of optical novelty filter based on bacteriorhodopsin film

    Institute of Scientific and Technical Information of China (English)

    Chen Gui-Ying; Yuan Yi-Zhe; Liang Xin; Xu Tang; Zhang Chun-Ping; Song Qi-Wang

    2006-01-01

    The quality of the novelty filter image is investigated at different intensities of the incident blue and yellow beams irradiating a bacteriorhodopsin (bR) film. The relationship between the transmitted blue beams and the incident yellow beams is established. The results show that the contrast of the novelty filter image depends on the lifetime of longest lived photochemical state (M state). These results enable one to identify the direction of a moving object and to improve the quality of the novel filter image by prolonging the lifetime of M state.

  17. Bacteriorhodopsin protein hybrids for chemical and biological sensing

    Science.gov (United States)

    Winder, Eric Michael

    Bacteriorhodopsin (bR), an optoelectric protein found in Halobacterium salinarum, has the potential for use in protein hybrid sensing systems. Bacteriorhodopsin has no intrinsic sensing properties, however molecular and chemical tools permit production of bR protein hybrids with transducing and sensing properties. As a proof of concept, a maltose binding protein-bacteriorhodopsin ([MBP]-bR) hybrid was developed. It was proposed that the energy associated with target molecule binding, maltose, to the hybrid sensor protein would provide a means to directly modulate the electrical output from the MBP-bR bio-nanosensor platform. The bR protein hybrid is produced by linkage between bR (principal component of purified purple membrane [PM]) and MBP, which was produced by use of a plasmid expression vector system in Escherichia coli and purified utilizing an amylose affinity column. These proteins were chemically linked using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), which facilitates formation of an amide bond between a primary carboxylic acid and a primary amine. The presence of novel protein hybrids after chemical linkage was analyzed by SDS-PAGE. Soluble proteins (MBP-only derivatives and unlinked MBP) were separated from insoluble proteins (PM derivatives and unlinked PM) using size exclusion chromatography. The putatively identified MBP-bR protein hybrid, in addition to unlinked bR, was collected. This sample was normalized for bR concentration to native PM and both were deposited onto indium tin oxide (ITO) coated glass slides by electrophoretic sedimentation. The photoresponse of both samples, activated using 100 Watt tungsten lamp at 10 cm distance, were equal at 175 mV. Testing of deposited PM with 1 mM sucrose or 1 mM maltose showed no change in the photoresponse of the material, however addition of 1 mM maltose to the deposited MBP-bR linked hybrid material elicited a 57% decrease in photoresponse

  18. Light-Induced Charge Separation and Transfer in Bacteriorhodopsin

    Institute of Scientific and Technical Information of China (English)

    HUANG Yu-Hua; LI Qing-Guo; ZHAO You-Yuan; ZHANG Zhong-Bin; OU-YANG Xiao-Ping; GONG Qin-Gan; CHEN Ling-Bing; LI Fu-Ming; LIU Jian; DING Jian-Dong

    2000-01-01

    The photo-voltage signals in bacteriorhodopsin(bR) excited by 1064nm pulse laser are different from those by 532 or 355 nm. It shows that the positive and negative photoelectric signals are produced by the motion of the positive and negative charges, respectively, and more energy is needed for producing the positive charges than the negative. The mechanism of light-induced charge generation and charge transfer in bR was studied and analyzed by measuring the photoelectric signals with different impedance of measuring circuit and different pulse-width of 532 nm laser as pump light.

  19. Engineering a Robust Photovoltaic Device with Quantum Dots and Bacteriorhodopsin

    OpenAIRE

    Renugopalakrishnan, Venkatesan; Barbiellini, Bernardo; King, Chris; Molinari, Michael; Mochalov, Konstantin; Sukhanova, Alyona; Nabiev, Igor; Fojan, Peter; Tuller, Harry L.; Chin, Michael; Somasundaran, Ponisseril; Padrós, Esteve; Ramakrishna, Seeram

    2014-01-01

    We present a route toward a radical improvement in solar cell efficiency using resonant energy transfer and sensitization of semiconductor metal oxides with a light-harvesting quantum dot (QD)/bacteriorhodopsin (bR) layer designed by protein engineering. The specific aims of our approach are (1) controlled engineering of highly ordered bR/QD complexes; (2) replacement of the liquid electrolyte by a thin layer of gold; (3) highly oriented deposition of bR/QD complexes on a gold layer; and (4) ...

  20. Effective photochromic nonlinearity of dried blue-membrane bacteriorhodopsin films

    Science.gov (United States)

    Tallent, Jack; Song, Q. Wang; Li, Zengfa; Stuart, Jeff; Birge, R. R.

    1996-09-01

    We report the effective nonlinearity for photochromic conversion in a blue-membrane bacteriorhodopsin film hosted in a dry polyvinyl alcohol matrix. The shift in absorption maximum on photoconversion in this film is larger than that of the same material in hydrated form, thus offering a larger modulation of the refractive index. The photoexcited index modulation is stable for several months, which provides for holographic data recording and long-term photochromic data storage. The effective index modulation is experimentally measured and is in good agreement with the theoretical predictions based on the Kramers-Kronig transformation.

  1. Spontaneous pattern formation in a thin film of bacteriorhodopsin with mixed absorptive-dispersive nonlinearity

    Science.gov (United States)

    Glückstad, J.; Saffman, M.

    1995-03-01

    We have observed the spontaneous formation of transverse spatial patterns in a thin film of bacteriorhodopsin with a feedback mirror. Bacteriorhodopsin has a mixed absorptive-dispersive nonlinearity at the wavelength used in the experiments (633 nm). Threshold values of the incident intensity for observation of pattern formation are found from a linear stability analysis of a model that describes bacteriorhodopsin as a sluggish saturable nonlinear medium with a complex Kerr coefficient. The calculated threshold intensity is in good agreement with the experimental observations, and the patterns are predicted to be frequency offset from the pump radiation.

  2. All-optical nonlinear holographic correlation using bacteriorhodopsin films

    Science.gov (United States)

    Thoma, Ralph; Dratz, Michael; Hampp, Norbert

    1995-05-01

    Films made of the halobacterial photochrome bacteriorhodopsin (BR) can be used in a number of holographic real-time applications. Their application as active material in a dual-axis joint- Fourier-transform (DAJFT) real-time correlator was shown recently. The BR films have a strong nonlinear intensity dependence on the light-induced absorption and refractive-index changes. Therefore the holographic diffraction efficiency also shows a nonlinear dependence on the writing intensity. We investigate the effect of this nonlinearity on the result of the correlation process in a bacteriorhodopsin-based DAJFT correlator. Numerical models supporting the experimental observations are presented. It was found that the BR film combines the holographic function for most objects with that of a spatial bandpass filter, whose center frequency is tuned by the writing intensity. This results in smaller peak widths and a suppression of the sidelobes. BR films allow the application of this nonlinear behavior in real time to the all-optical correlation process.

  3. Detecting conformational change by current transport in Bacteriorhodopsin

    CERN Document Server

    Alfinito, Eleonora

    2008-01-01

    Charge transport modification exhibiting an increase of conductance activated by an external green light in Bacteriorhodopsin is correlated to its conformational change. A theoretical model based on a map of the protein structure into a resistor network is implemented to account for a sequential tunneling mechanism of charge transfer through neighbouring amino-acids. The model is validated by comparison with current-voltage experiments and provides for the potential barriers involved in the charge transfer an average height of 69 meV over an interacting radius of 6 \\aa. The predictability of the model is also tested on bovine rhodopsin, the prototype of the G protein coupled receptor (GPCR) family also sensitive to the light, with results exhibiting the opposite behaviour of a decrease of conductance in the presence of light.

  4. Thermochromism of bacteriorhodopsin and its pH dependence.

    Science.gov (United States)

    Neebe, Martin; Rhinow, Daniel; Schromczyk, Nina; Hampp, Norbert A

    2008-06-12

    Purple membranes (PMs), which consist of the photochromic membrane protein bacteriorhodopsin (BR) and lipids only, show complex thermochromic properties. Three different types of reversible temperature-dependent spectral transitions were found, involving spectral states absorbing at 460, 519, and 630 nm. These thermochromic absorption changes were analyzed in the range from 10 to 80 degrees C. In dependence on the bulk pH value, hypsochromic or bathochromic shifts in the BR absorption spectra are observed in BR gels as well as in BR films. The thermochromic changes between both purple and blue or purple and red were quantified in the CIE color system. The molecular changes causing these effects are discussed, and a model is presented in terms of intramolecular protonation equilibriums. The thermochromic properties of BR may be of interest in applications like security tags, as this feature may complement the well-known photochromic properties of BR. PMID:18491932

  5. Bacteriorhodopsin films for optical signal processing and data storage

    Science.gov (United States)

    Walkup, John F. (Principal Investigator); Mehrl, David J. (Principal Investigator)

    1996-01-01

    This report summarizes the research results obtained on NASA Ames Grant NAG 2-878 entitled 'Investigations of Bacteriorhodopsin Films for Optical Signal Processing and Data Storage.' Specifically we performed research, at Texas Tech University, on applications of Bacteriorhodopisin film to both (1) dynamic spatial filtering and (2) holographic data storage. In addition, measurements of the noise properties of an acousto-optical matrix-vestor multiplier built for NASA Ames by Photonic Systems Inc. were performed at NASA Ames' Photonics Laboratory. This research resulted in two papers presented at major optical data processing conferences and a journal paper which is to appear in APPLIED OPTICS. A new proposal for additional BR research has recently been submitted to NASA Ames Research Center.

  6. Bacteriorhodopsin-based bipolar photosensor for biomimetic sensing

    Science.gov (United States)

    Kasai, Katsuyuki; Haruyama, Yoshihiro; Yamada, Toshiki; Akiba, Makoto; Tominari, Yukihiro; Kaji, Takahiro; Terui, Toshifumi; Peper, Ferdinand; Tanaka, Shukichi; Katagiri, Yoshitada; Kikuchi, Hiroshi; Okada-Shudo, Yoshiko; Otomo, Akira

    2013-10-01

    Bacteriorhodopsin (bR) is a promising biomaterial for several applications. Optical excitation of bR at an electrode-electrolyte interface generates differential photocurrents while an incident light is turned on and off. This unique functional response is similar to that seen in retinal neurons. The bR-based bipolar photosensor consists of the bR dip-coated thin films patterned on two ITO plates and the electrolyte solution. This bipolar photocell will function as a biomimetic photoreceptor cell. The bipolar structure, due to the photocurrent being generated in alignment with the cathodic direction, makes the excitatory and inhibitory regions possible. This scheme shows our bipolar cell can act as a basic unit of edge detection and forms the artificial visual receptive field.

  7. Integrated optical switching based on the protein bacteriorhodopsin.

    Science.gov (United States)

    Dér, András; Valkai, Sándor; Fábián, László; Ormos, Pál; Ramsden, Jeremy J; Wolff, Elmar K

    2007-01-01

    According to our earlier pioneering study, a dry film containing native bacteriorhodopsin (bR) shows unique nonlinear optical properties (refractive index change, controllable by light of different colors, greater than 2 x 10(-3)) that are in many respects superior to those of the materials presently applied in integrated optics. Here, we report on the first integrated optical application based on a miniature Mach-Zehnder interferometer (see Figs. 1 and 2) demonstrating a real switching effect by bR (efficiency higher than 90%) due to the M-state. Our results also imply that the refractive index change of the K-state (9 x 10(-4)) is high enough for fast switching. PMID:17132043

  8. Generalized model for all-optical light modulation in bacteriorhodopsin

    Science.gov (United States)

    Roy, Sukhdev; Singh, C. P.; Reddy, K. P. J.

    2001-10-01

    We present a generalized model for the photochemical cycle of bacteriorhodopsin (bR) protein molecule. Rate equations have been solved for the detailed light-induced processes in bR for its nine states: B→K↔L↔MI→MII↔N↔O↔P→Q→B. The complete steady-state intensity-induced population densities in various states of the molecule have been computed to obtain a general, exact, and analytical expression for the nonlinear absorption coefficient for multiple modulation pump laser beams. All-optical light modulation of different probe laser beam transmissions by intensity induced population changes due to one and two modulation laser beams has been analyzed. The proposed model has been shown to accurately model experimental results.

  9. Bacteriorhodopsin-the basis of molecular superfast nanoelectronics

    Science.gov (United States)

    Samoilovich, M. I.; Belyanin, A. F.; Grebennikov, E. P.; Guriyanov, A. V.

    2002-12-01

    We give some perspectives on the possibilities for application of the bacteriorhodopsin (BR) molecule in the development of nanoelectronic devices in conjunction with photonic crystals-materials with inhibited photonic band. This involves the use of a single BR molecule by `changing' the photon gradient formation with externally initiated electric fields, preserving the mechanisms of negative reverse communication, or the use of an intermolecular mechanism of photo transitions and dependence on the electric field. Greatly enhanced optical responses, linear and nonlinear, in metal nanocomposites and nanodiamond containing nanoscale surface features have been intensively studied in recent years. These results foreshadow fascinating possibilities for linear and nonlinear local spectroscopy of single molecules. Use of photonic crystals (at present the only technologically realized three-dimensional photonic crystals are the opal matrices) suggests the possible application of a number of physical phenomena.

  10. High-contrast, all-optical switching in bacteriorhodopsin films

    Science.gov (United States)

    Banyal, Ravinder Kumar; Raghavendra Prasad, B.

    2005-09-01

    We report experiments with nonlinear-absorption-based, high-contrast, all-optical switching in photochromic bacteriorhodopsin (BR) films. The switching action is accomplished by control of the transmission of a weak probe beam through a BR sample with the help of strong pump beam illumination at 532 nm wavelength. We found that the switching properties of BR films depend on several experimentally controllable parameters such as probe wavelength, pump beam intensity, and excitation rate. A comparative study of the switching behavior and other parameters of practical use was carried out at three probe wavelengths (543, 594, and 633 nm) and various beam powers and pump excitation rates. The results are presented for commercially available wild-type and D96N variant BR films.

  11. Nonlinear Optical Interactions in Bacteriorhodopsin Using Z-Scan

    Science.gov (United States)

    Aranda, Francisco J.; Rao, Devulapalli V. G. L. N.; Wong, Chi L.; Zhou, Ping; Chen, Zhong; Akkara, Joseph A.; Kaplan, David L.; Roach, Joseph F.

    1995-06-01

    Nonlinear refractive index coefficient n2 of bacteriorhodopsin suspensions in water is measured by the Z-scan technique with a low power continuous wave laser at 647.1 manometer wavelength. Our results indicate that both the magnitude and the sign of n2 depend strongly on the light intensity. Negative values for n2 are obtained for on axis laser irradiance at the focus above 3 W/cm2. The observed self-defocusing phenomena can be attributed to the index change due to the light induced transition between the photochromic states. The results elucidate the origin of n2 and offer a plausible explanation for the differences in the reported n2 measurements.

  12. Application of Bacteriorhodopsin Films in an Adaptive-Focusing Schlieren System

    Science.gov (United States)

    Downie, John D.

    1995-01-01

    The photochromic property of bacteriorhodopsin films is exploited in the application of a focusing schlieren optical system for the visualization of optical phase information. By encoding an image on the film with light of one wavelength and reading out with a different wavelength, the readout beam can effectively see the photographic negative of the original image. The potential advantage of this system over previous focusing schlieren systems is that the updatable nature of the bacteriorhodopsin film allows system adaptation. I discuss two image encoding and readout techniques for the bacteriorhodopsin and use film transmission characteristics to choose the more appropriate method. I demonstrate the system principle with experimental results using argon-ion and He-Cd lasers as the two light sources of different wavelengths, and I discuss current limitations to implementation with a white-light source.

  13. Dynamic Behaviour of Self-Diffraction in Bacteriorhodopsin Film

    Institute of Scientific and Technical Information of China (English)

    GUO Zong-Xia; CHEN Gui-Ying; ZHANG Chun-Ping; TIAN Jian-Guo; Q. Wang Song; SHEN Bin; FU Guang-Hua

    2004-01-01

    @@ We investigate the dependences of the diffraction efficiency of the first order self-diffracted beam in bacteriorhodopsin (bR) films on the illumination time, the intensity and wavelength of the incident light. When the blue light (λ = 488 nm) and low intensity red light (λ = 632.8 nm) are incident on the bR film respectively,the diffraction efficiencies increase from zero to a stable value with the illumination time. When the green light (λ = 533 nm) and high-intensity red light illuminate the bR film respectively, the diffraction efficiencies increase from zero to the maximum and then decrease to a stable value with the illumination time. Rise and decay times are dependent on the intensity and wavelength of the incident light. The maximaldiffraction efficiency of the red light is twice as high as that of the green light. The highest diffraction efficiency of 5.4% is obtained at 633nm.The diffraction efficiency change with the time for the green light is larger than that for the blue and red light.

  14. Engineering a Robust Photovoltaic Device with Quantum Dots and Bacteriorhodopsin.

    Science.gov (United States)

    Renugopalakrishnan, Venkatesan; Barbiellini, Bernardo; King, Chris; Molinari, Michael; Mochalov, Konstantin; Sukhanova, Alyona; Nabiev, Igor; Fojan, Peter; Tuller, Harry L; Chin, Michael; Somasundaran, Ponisseril; Padrós, Esteve; Ramakrishna, Seeram

    2014-07-31

    We present a route toward a radical improvement in solar cell efficiency using resonant energy transfer and sensitization of semiconductor metal oxides with a light-harvesting quantum dot (QD)/bacteriorhodopsin (bR) layer designed by protein engineering. The specific aims of our approach are (1) controlled engineering of highly ordered bR/QD complexes; (2) replacement of the liquid electrolyte by a thin layer of gold; (3) highly oriented deposition of bR/QD complexes on a gold layer; and (4) use of the Forster resonance energy transfer coupling between bR and QDs to achieve an efficient absorbing layer for dye-sensitized solar cells. This proposed approach is based on the unique optical characteristics of QDs, on the photovoltaic properties of bR, and on state-of-the-art nanobioengineering technologies. It permits spatial and optical coupling together with control of hybrid material components on the bionanoscale. This method paves the way to the development of the solid-state photovoltaic device with the efficiency increased to practical levels. PMID:25383133

  15. Integrated optical devices using bacteriorhodopsin as active nonlinear optical material

    Science.gov (United States)

    Dér, András; Fábián, László; Valkai, Sándor; Wolff, Elmar; Ramsden, Jeremy; Ormos, Pál

    2006-08-01

    Coupling of optical data-processing devices with microelectronics, telecocommunication and sensory functions, is among the biggest challenges in molecular electronics. Intensive research is going on to find suitable nonlinear optical materials that could meet the demanding requirements of optoelectronic applications, especially regarding high sensitivity and stability. In addition to inorganic and organic crystals, biological molecules have also been considered for use in integrated optics, among which the bacterial chromoprotein, bacteriorhodopsin (bR) generated the most interest. bR undergoes enormous absorption and concomitant refractive index changes upon initiation of a cyclic series of photoreactions by a burst of actinic light. This effect can be exploited to create highly versatile all-optical logical elements. We demonstrate the potential of this approach by investigating the static and dynamic response of several basic elements of integrated optical devices. Our results show that, due to its relatively high refractive index changes, bR can be used as an active nonlinear optical material to produce a variety of integrated optical switching and modulation effects.

  16. All-optical biomolecular parallel logic gates with bacteriorhodopsin.

    Science.gov (United States)

    Sharma, Parag; Roy, Sukhdev

    2004-06-01

    All-optical two input parallel logic gates with bacteriorhodopsin (BR) protein have been designed based on nonlinear intensity-induced excited-state absorption. Amplitude modulation of a continuous wave (CW) probe laser beam transmission at 640 nm corresponding to the peak absorption of O intermediate state through BR, by a modulating CW pump laser beam at 570 nm corresponding to the peak absorption of initial BR state has been analyzed considering all six intermediate states in its photocycle using the rate equation approach. The transmission characteristics have been shown to exhibit a dip, which is sensitive to normalized small-signal absorption coefficient (beta), rate constants of O and N intermediate states and absorption of the O state at 570 nm. There is an optimum value of beta for a given pump intensity range for which maximum modulation can be achieved. It is shown that 100% modulation can be achieved if the initial state of BR does not absorb the probe beam. The results have been used to design low-power all-optical parallel NOT, AND, OR, XNOR, and the universal NAND and NOR logic gates for two cases: 1) only changing the output threshold and 2) considering a common threshold with different beta values. PMID:15382746

  17. Alternative translocation of protons and halide ions by bacteriorhodopsin

    International Nuclear Information System (INIS)

    Bacteriorhodopsin (bR568) in purple membrane near pH 2 shifts its absorption maximum from 568 to 605 nm forming the blue protein bRacid605, which no longer transports protons and which shows no transient deprotonation of the Schiff base upon illumination. Continued acid titration with HCl or HBr but not H2SO4 restores the purple chromophore to yield bRHCl564 or bRHBr568. These acid purple forms also regain transmembrane charge transport, but no transient Schiff base deprotonation is observed. In contrast to bR568, no rate decrease of the bRacidpurple transport kinetics is detected in 2H2O; however, the transport rate decreases by a factor of ∼ 2 in bRHBr568 compared with bRHCl564. The data indicate that in the acid purple form bR transports the halide anions instead of protons. The authors present a testable model for the transport mechanism, which should also be applicable to halorhodopsin

  18. Electron microscopic observation and rotational diffusion measurement of bacteriorhodopsin in lipid vesicles

    Institute of Scientific and Technical Information of China (English)

    HU; Kunsheng

    2001-01-01

    [1]Stoeckenius, W.. Bacterial rhodopsins: Evolution of a mechanistic model for the ion pumps, Protein Science, 1999, 8: 447.[2]Ebrey. T. G, Light energy transduction in bacteriorhodopsin, in Thermodynamics of Membranes, Receptors and Channels (ed. Jackson. M.), New York: CRC Press, 1993. 353-387.[3]Lanyi. J. K.. Understanding structure and function in the light-driven proton pump bacteriorhodopsin, J. Struct. Biol., 1998,l24: 164.[4]Quay. S. C., Condie. C. C., Conformational studies of aqueous melittin: Thermodynamic parameters of the monomer-tetramer self-association reaction. Biochemistry, 1983, 22: 695.[5]Habermann. E.. Bee and wasp venoms. Science, 1972, 177: 314.[6]Tosteson. M. T., Holmes. S. J., Razin. M. et al., Melitton lysis of red cells, J. Membr. Biol., 1985, 87: 35.[7]Hu, K. S., Dufton, M. J., Morrison, I. E. G. et al., Cherry interaction of bee venom melittin with bacteriorhodopsin in lipid vesicles: Protein rotational diffusion measurement. Biochem. Biophys. Acta, 1985, 816(2): 358.[8]Shi, H., Hu, K. S., Huang, Y. et al., Effect of melittin on photocycle and photoresponse of purple membrane: sites of interaction between bacteriorhodopsin and melittin, Photochemistry and Photobiology, 1993, 58(3): 413.[9]Jiang. Q. X., Hu, K. S.. Shi. H., Interaction of both melittin and its site-specific mutants with bacteriorhodopsin of Halobacterium halobium: sites of electrostatic interaction on melittin Photochemistry and Photobiology, 1994, 60(2): 175.[10]Doebler, R., Basaran. N.. Goldston H. et al., Effect of protein aggregation into aqueous phase on the binding of membrane proteins to membranes, Biophys. J., 1999, 76: 928.[11]Rehorek, M., Heyn, M. P, Binding of all-trans-retinal to the purple membrane, Evidence for cooperativity and determination of the extinction coefficient, Biochemistry, 1979, 18: 4977.[12]Chen. P. S. Jr.. Toribara, T. Y., Warner, H., Microdetermination of phosphorous, Anal. Chem., t956, 28

  19. Nonlinear Coherent Optical Image Processing Using Logarithmic Transmittance of Bacteriorhodopsin Films

    Science.gov (United States)

    Downie, John D.

    1995-01-01

    The transmission properties of some bacteriorhodopsin-film spatial light modulators are uniquely suited to allow nonlinear optical image-processing operations to be applied to images with multiplicative noise characteristics. A logarithmic amplitude-transmission characteristic of the film permits the conversion of multiplicative noise to additive noise, which may then be linearly filtered out in the Fourier plane of the transformed image. I present experimental results demonstrating the principle and the capability for several different image and noise situations, including deterministic noise and speckle. The bacteriorhodopsin film studied here displays the logarithmic transmission response for write intensities spanning a dynamic range greater than 2 orders of magnitude.

  20. Nonlinear coherent optical image processing using logarithmic transmittance of bacteriorhodopsin films

    Science.gov (United States)

    Downie, John D.

    1995-08-01

    The transmission properties of some bacteriorhodopsin-film spatial light modulators are uniquely suited to allow nonlinear optical image-processing operations to be applied to images with multiplicative noise characteristics. A logarithmic amplitude-transmission characteristic of the film permits the conversion of multiplicative noise to additive noise, which may then be linearly filtered out in the Fourier plane of the transformed image. I present experimental results demonstrating the principle and the capability for several different image and noise situations, including deterministic noise and speckle. The bacteriorhodopsin film studied here displays the logarithmic transmission response for write intensities spanning a dynamic range greater than 2 orders of magnitude.

  1. Spatial light modulation based on photoinduced change in the complex refractive index of bacteriorhodopsin

    Science.gov (United States)

    Takei, Hiroyuki; Shimizu, Norio

    1996-04-01

    Bacteriorhodopsin exhibits photoinduced changes in both absorption and refractive index at 633 nm. To explore the possibility of exploiting this property in constructing a photoaddressed spatial light modulator, we investigated the transmission property of a Fabry-Perot interferometer containing a bacteriorhodopsin thin film. Film was formed that had a phase shift of pi /4 and sufficient interference fringe contrast for spatial light modulation. This establishes the possibility of constructing a spatial light modulator that features nonlinear input-output characteristics and can operate at moderate light intensities of the order of tens of milliwatts per centimeter square. spatial light modulation, complex refractive index.

  2. Part I. Bacteriorhodopsin-related materials work for molecular electronics. Part II. Volumetric optical memory based on the branched photocycle of bacteriorhodopsin. Part III. The role of calcium in the bacteriorhodopsin binding site

    Science.gov (United States)

    Stuart, Jeffrey Alan

    Part I. A protocol for the routine isolation and purification of purple membrane sheets containing the integral membrane protein, bacteriorhodopsin, was developed based upon modifications of protocols already in the literature. This simplified protocol is geared toward the facile isolation of protein for use in molecular electronic devices. Methods for the incorporation of bacteriorhodopsin into various polymeric supports were also developed, primarily in the form of dried films and hydrated cubes. This work also represents the first reported production of dried films of the deionized protein, or blue membrane. Part II. An architecture for a volumetric optical memory based on the branched-photocycle of bacteriorhodopsin is presented. The branching reaction circumvents problems associated with destructive reading and writing processes and allows access to a stable, long-lived state, separated both temporally and energetically from the main photocycle, thereby making long-term data storage possible. The state, denoted as Q, can only be accessed by exposing the protein to two different wavelengths of light in the proper sequence, with the appropriate temporal separation (roughly 2 ms between the light pulses). The Q-state (assigned as a binary one) is transparent to both writing and reading processes, making them rigorously non-destructive. Bacteriorhodopsin in its resting state is assigned as a binary zero. A differential absorption reading process is used to determine the state of each volumetric binary element. Preliminary results are reported. Part III. The nature of the chromophore binding site of light-adapted bacteriorhodopsin is analyzed by using all-valence electron MNDO and MNDO-PSDCI molecular orbital theory to interpret previously reported linear and nonlinear optical spectroscopic measurements. It is concluded that the unique two-photon properties of the chromophore are due in part to the electrostatic field associated with a Casp{2+} ion near the

  3. White Light Schlieren Optics Using Bacteriorhodopsin as an Adaptive Image Grid

    Science.gov (United States)

    Peale, Robert; Ruffin, Boh; Donahue, Jeff; Barrett, Carolyn

    1996-01-01

    A Schlieren apparatus using a bacteriorhodopsin film as an adaptive image grid with white light illumination is demonstrated for the first time. The time dependent spectral properties of the film are characterized. Potential applications include a single-ended Schlieren system for leak detection.

  4. Study of nonlinear optical properties of multilayer Langmuir-Blodgett films containing bacteriorhodopsin.

    Science.gov (United States)

    Barmenkov Yu, O; Kir'yanov, A V; Starodumov, A N; Maslyanitsyn, I A; Shigorin, V D; Lemmetyinen, H

    2000-08-01

    Multilayer oriented Langmuir-Blodgett films of bacteriorhodopsin were prepared and their nonlinear optical properties, including second harmonic generation and photoresponse at a two phase-modulated beams mixing, were investigated. The nonlinear component of refractive index of the films was measured. PMID:10946566

  5. Real-time self-induced nonlinear optical Zernike-type filter in a bacteriorhodopsin film

    Science.gov (United States)

    Iturbe Castillo, David; Sanchez-de-la-Llave, David; Garcia, Ruben R.; Olivos-Perez, L. I.; Gonzalez, Luis A.; Rodriguez-Ortiz, M.

    2001-11-01

    We propose the use of a nonlinear bacteriorhodopsin film to self-induce a Zernike-type filter in robust optical phase-contrast systems. The device requires relatively low light intensity levels (as low as 200 nW/cm2) at wavelengths around 633 nm and can contrast dynamical phase distributions.

  6. Determination of the thermal expansion and thermo-optic coefficients of a bacteriorhodopsin film

    Science.gov (United States)

    Wang Song, Q.; Zhang, Chunping; Ku, Chin-Yu; Huang, Ming-Chieh; Gross, Richard B.; Birge, Robert R.

    1995-02-01

    The linear expansion and thermo-optic coefficients of a bacteriorhodopsin film were measured by using an interferometric method. The experimental results confirm the previous suspicions that the large refractive nonlinearity which occurs at high illumination intensities arises form a thermal effect. The results also suggest a possible way to increase the usable thermal nonlinearity by four times.

  7. Optical switching in bistable active cavity containing nonlinear absorber on bacteriorhodopsin

    Science.gov (United States)

    Bazhenov, Vladimir Y.; Taranenko, Victor B.; Vasnetsov, Mikhail V.

    1993-04-01

    The transverse nonlinear dynamics of switchings in an active system (laser with nonlinear saturable absorber on bacteriorhodopsin in a self-imaging cavity) is studied both experimentally and theoretically. The soliton-like light field structure formation and continuously cycled self-switching process are investigated.

  8. Evidence of multipolar response of Bacteriorhodopsin by noncollinear second harmonic generation.

    Science.gov (United States)

    Bovino, F A; Larciprete, M C; Sibilia, C; Váró, G; Gergely, C

    2012-06-18

    Noncollinear second harmonic generation from a Bacteriorhodopsin (BR) oriented multilayer film was systematically investigated by varying the polarization state of both fundamental beams. Both experimental results and theoretical simulations, show that the resulting polarization mapping is an useful tool to put in evidence the optical chirality of the investigated film as well as the corresponding multipolar contributions to the nonlinear. PMID:22714524

  9. Trapping the M sub 1 and M sub 2 substrates of bacteriorhodopsin for electron diffraction studies

    Energy Technology Data Exchange (ETDEWEB)

    Perkins, G.A.

    1992-05-01

    Visible and Fourier transform infrared (FTIR) absorption spectroscopies are used to observe protein conformational changes occuring during the bacteriorhodopsin photocycle. Spectroscopic measurements which define the conditions under which bacteriorhodopsin can be isolated and trapped in two distinct substates of the m intermediate of the photocycle, M{sub 1}, and M{sub 2}, are described. A protocol that can be used for high-resolution electron diffraction studies is presented that will trap glucose-embedded purple membrane in the M{sub 1}and M{sub 2} substates at greater than 90% concentration. It was discovered that glucose alone does not provide a fully hydrated environment for bacteriorhodopsin. Equilibration of glucose-embedded samples at high humidity can result in a physical state that is demonstrably closer to the native, fully hydrated state. An extension of the C-T Model of bacteriorhodopsin functionality (Fodor et al., 1988; Mathies et al., 1991) is proposed based on FTIR results and guided by published spectra from resonance Raman and FTIR work. 105 refs.

  10. Time-resolved laser studies on the proton pump mechanism of bacteriorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    El-Sayed, M.A.

    1991-01-01

    The research work carried out in 1991 can be classified as follows: (1) Work on the nature of the binding site of Eu{sup 3+} in which a fluorescence technique was used to determine the binding equilibrium constant from the concentration of the free Eu{sup 3+} in equilibrium with the bound ions. (2) The mechanism of the slow deprotonation process of bacteriorhodopsin during its photocycle from the observed temperature and pH dependence of its kinetics. (3) Using the circular dichroism spectrum of bR and its perturbed forms to examine the nature of the primary process as well as the origin of the non-exponential kinetic behavior of its photocycle. (4) Studies of bacteriorhodopsin mutants to identify the important amino acids that are part of the reaction coordinate of the deprotonation process as well as to assign the species that are important in giving rise to UV transient absorption whose origin was controversial.

  11. Metal ion binding sites of bacteriorhodopsin. Laser-induced lanthanide luminescence study

    International Nuclear Information System (INIS)

    Laser-excited luminescence lifetimes of lanthanide ions bound to bacteriorhodopsin have been measured in deionized membranes. The luminescence titration curve, as well as the binding curve of apomembrane (retinal-free) with Eu3+, has shown that the removal of the retinal does not significantly affect the affinity of Eu3+ for the two high affinity sites of bacteriorhodopsin. The D2O effects on decay rate constants indicate that Eu3+ bound to the high affinity sites of native membrane or apomembrane is coordinated by about six ligands in the first coordination sphere. Tb3+ is shown to be coordinated by four ligands. The data indicate that metal ions bind to the protein with a specific geometry. From intermetal energy transfer experiments using Eu3+-Pr3+, Tb3+-Ho3+, and Tb3+-Er3+, the distance between the two high affinity sites is estimated to be 7-8 A

  12. Tryptophan fluorescence quenching by alkaline earth metal cations in deionized bacteriorhodopsin.

    Science.gov (United States)

    Wang, G; Wang, A J; Hu, K S

    2000-12-01

    Tryptophan quenching by the addition of alkaline earth metal cations to deionized bacteriorhodopsin suspensions was determined. The results show that the addition of cation primarily quenches fluorescence from surface tryptophan residues. The quenched intensity exhibits a 1/R dependence, where R is the ionic radius of the corresponding metal ion. This observation results from a stronger energy transfer coupling between the tryptophan and the retinal. The membrane curvature may be involved as a result of cations motion and correlated conformational changes. PMID:11332888

  13. Optical signal processing on photorefractive crystal substrate and bacteriorhodopsin thin film

    Science.gov (United States)

    Azimipour, Mehdi; Pashaie, Ramin

    2013-03-01

    In this article we present a new approach for implementation of computation algorithms to perform nonlinear signal processing with light on the surface of a photorefractive crystal and Bacteriorhodopsin thin film. Using the developed mathematical models for the photodynamics of these materials, we demonstrate a specific operation mode and a design procedure to obtain nonlinear response which can be used for implementation of high-performance photonic computers.

  14. Polarization properties of four-wave interaction in dynamic recording material based on bacteriorhodopsin

    Science.gov (United States)

    Korchemskaya, Ellen Y.; Soskin, Marat S.

    1994-10-01

    The polarization properties of four-wave interaction on polymer films with bacteriorhodopsin that possess anisotropically saturating nonlinearity are studied both theoretically and experimentally. The amplitude and the polarization of the diffracted wave for recording material with anisotropically saturating nonlinearity are calculated. Low saturation intensity allows the operation of the polarization of low-intensity signals to be realized. It is shown that control of the diffractive wave polarization is possible only with the variation of the light recording intensity.

  15. The 3rd-order nonlinearity of bacteriorhodopsin by four-wave mixing

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The 3rd-order nonlinear optical susceptibility X(3) and the response time of the light-transducing biomolecule bacteriorhodopsin were measured with the four-wave mixing technique and a picosecond frequency-doubled Nd:YAG laser(532nm).The X(3) and the response time measured are 10-9 esu and 20 ps,respectively.The possible mechanism for generating the 3rd-order nonlinear optical susceptibility X(3) and response time were discussed.

  16. Optical chirality of bacteriorhodopsin films via second harmonic Maker's fringes measurements

    Science.gov (United States)

    Larciprete, M. C.; Belardini, A.; Sibilia, C.; Saab, M.-b.; Váró, G.; Gergely, C.

    2010-05-01

    We experimentally investigated second harmonic generation from an oriented multilayer film of bacteriorhodopsin protein, deposited onto a charged surface. The generated signal is obtained as a function of incidence angle, at different polarization state of both fundamental and generated beams. We show that the measurements, together with the analytical curves, allow to retrieve the nonvanishing elements of the nonlinear optical tensor, including the ones introduced by optical chirality.

  17. Light-induced changes in the absorption spectrum of bacteriorhodopsin under two-wavelength excitation

    Science.gov (United States)

    Koklyushkin, A. V.; Korolev, A. E.

    2004-09-01

    The results of spectrophotometric measurements of nonlinear light-induced changes in the absorption spectrum of bacteriorhodopsin D96N occurring upon simultaneous excitation at the wavelengths 633 and 441 nm in the excitation intensity range typical for recording of dynamic holograms are presented. The quantitative conditions under which the action of the radiation at one wavelength reduces the change in the optical density caused by the radiation at the other wavelength are determined.

  18. Can the Low-Resolution Structures of Photointermediates of Bacteriorhodopsin Explain Their Crystal Structures?

    OpenAIRE

    Kamikubo, Hironari; Kataoka, Mikio

    2004-01-01

    To understand the molecular mechanism of light-driven proton pumps, the structures of the photointermediates of bacteriorhodopsin have been intensively investigated. Low-resolution diffraction techniques have demonstrated substantial conformational changes at the helix level in the M and N intermediates, between which there are noticeable differences. The intermediate structures at atomic resolution have also been solved by x-ray crystallography. Although the crystal structures have demonstra...

  19. Two-photon polarization data storage in bacteriorhodopsin films and its potential use in security applications

    International Nuclear Information System (INIS)

    Bacteriorhodopsin (BR) films allow write-once-read-many recording of polarization data by a two-photon-absorption (TPA) process. The optical changes in BR films induced by the TPA recording were measured and the Müller matrix of a BR film was determined. A potential application of BR films in security technology is shown. Polarization data can be angle-selective retrieved with high signal-to-noise ratio. The BR film does not only carry optical information but serves also as a linear polarizer. This enables that polarization features recorded in BR films may be retrieved by merely using polarized light from a mobile phone display

  20. Photoinduced Birefringence and Broadband All-Optical Photonic Switch in a Bacteriorhodopsin/Polymer Composite Film

    Institute of Scientific and Technical Information of China (English)

    WEI Lai; TENG Xue-Lei; Lu Ming; ZHAO You-Yuan; MA De-Wang; DING Jian-Dong

    2007-01-01

    Photoinduced birefringence with large optical nonlinearity in a bacteriorhodopsin/polymer composite film is observed.A high refractive index change of 8.5×10-5 photoinduced by 476nm pumping beam is reached at the low intensity of 6.5mW/cm2.Based on it,a broadband all-optical photonic switch is realized with an optical controlling switch system.Because of controlling beam's selectivity in switching,the transporting beams of different wavelengths with different intensities and shapes can be modulated by adjusting the wavelength and intensity of the controlling beam.

  1. Nonlinear photoinduced anisotropy and modifiable optical image display in a bacteriorhodopsin/polymer composite film

    Science.gov (United States)

    Wei, Lai; Luo, Jia; Zhu, Jiang; Lu, Ming; Zhao, You-yuan; Ma, De-wang; Ding, Jian-dong

    2007-04-01

    The nonlinear photoinduced anisotropy with large birefringence in a bacteriorhodopsin/polymer composite (bR/PC) film was observed. The contrast ratio, a ratio of the maximum to the minimum intensity of transmitted probe light through the bR/PC film within the linear gray scale range could reach ˜350:1. An all-optical image display in different colors was performed. The intensity of the transmitted signal could be modulated by adjusting the multibeam polarization states and intensities. Therefore, the positive image, negative image, and image erasure in display were demonstrated.

  2. A Novel Optical Filter for Removing Bright-Background Using an Enhanced Bacteriorhodopsin Thin Film

    Institute of Scientific and Technical Information of China (English)

    ZHANG Chun-Ping(张春平); GU Li-Qun(顾立群); CHEN Gm-Ying(陈桂英); GUO Zong-Xia(郭宗霞); FU Guang-Hua(富光华); ZHANG Guang-Yin(张光寅); ZHANG Tian-Hao(张天浩); LI Yu-Dong(李玉栋); Q. Wang SONG

    2003-01-01

    We found that the bacteriorhodopsin (bR) film has a special property of complementary suppression modulated transmission (CSMT). The yellow and the blue beams can be suppressed mutually when both the beams illuminate the bR film simultaneously. When the blue beam carrying an image with a bright-background noise illuminates on the bR film and then a yellow beam with uniform intensity distribution illuminates the same area, the brightbackground can be removed due to the CSMT. In our demonstration, the pattern model is letters "VLSI" with ground noise of small words and the ground noise is removed from the pattern by the new optical filter.

  3. Förster Resonance Energy Transfer between Core/Shell Quantum Dots and Bacteriorhodopsin

    OpenAIRE

    Karna, Shashi P.; Craig R. Friedrich; Garrett, Gregory A.; Lueking, Donald R.; Winder, Eric M.; Griep, Mark H.

    2012-01-01

    An energy transfer relationship between core-shell CdSe/ZnS quantum dots (QDs) and the optical protein bacteriorhodopsin (bR) is shown, demonstrating a distance-dependent energy transfer with 88.2% and 51.1% of the QD energy being transferred to the bR monomer at separation distances of 3.5 nm and 8.5 nm, respectively. Fluorescence lifetime measurements isolate nonradiative energy transfer, other than optical absorptive mechanisms, with the effective QD excited state lifetime reducing from 18...

  4. General model for lipid-mediated two-dimensional array formation of membrane proteins: Application to bacteriorhodopsin

    DEFF Research Database (Denmark)

    Sabra, Mads Christian; Uitdehaag, J.C.M.; Watts, A

    1998-01-01

    Based on experimental evidence for 2D array formation of bacteriorhodopsin, we propose a general model for lipid-mediated 2D array formation of membrane proteins in lipid bilayers. The model includes two different lipid Species; "annular" lipids and "neutral" lipids, and one protein species. The ...

  5. Contrast enhancement and phase conjugation low-power optical signal in dynamic recording material based on bacteriorhodopsin

    Science.gov (United States)

    Korchemskaya, Elena Y.; Soskin, Marat S.; Dukova, Tatjana V.; Vsevolodov, Nicolai N.

    1994-02-01

    Polymer films with fragments of the purple membranes containing protein bacteriorhodopsin (BR) have been used for the real-time optical information processing of low-power (several milliwatt) cw gas laser signals. The nonlinear recording media with BR have a potential in microscopic techniques for in-vivo diagnosis of the crystalline lens.

  6. Trapping the M{sub 1} and M{sub 2} substrates of bacteriorhodopsin for electron diffraction studies

    Energy Technology Data Exchange (ETDEWEB)

    Perkins, G.A.

    1992-05-01

    Visible and Fourier transform infrared (FTIR) absorption spectroscopies are used to observe protein conformational changes occuring during the bacteriorhodopsin photocycle. Spectroscopic measurements which define the conditions under which bacteriorhodopsin can be isolated and trapped in two distinct substates of the m intermediate of the photocycle, M{sub 1}, and M{sub 2}, are described. A protocol that can be used for high-resolution electron diffraction studies is presented that will trap glucose-embedded purple membrane in the M{sub 1}and M{sub 2} substates at greater than 90% concentration. It was discovered that glucose alone does not provide a fully hydrated environment for bacteriorhodopsin. Equilibration of glucose-embedded samples at high humidity can result in a physical state that is demonstrably closer to the native, fully hydrated state. An extension of the C-T Model of bacteriorhodopsin functionality (Fodor et al., 1988; Mathies et al., 1991) is proposed based on FTIR results and guided by published spectra from resonance Raman and FTIR work. 105 refs.

  7. Studying of Phototransformation of Light Signal by Photoreceptor Pigments - Rhodopsin, Iodopsin and Bacteriorhodopsin

    Directory of Open Access Journals (Sweden)

    Ignat Ignatov

    2014-09-01

    Full Text Available This review article views predominately the structure and function of animal and bacterial photoreceptor pigments (rhodopsin, iodopsin, bacteriorhodopsin and their aspects of nano- and biotechnological usage. On an example of bacteriorhodopsin is described the method of its isolation from purple membranes of photo-organotrophic halobacterium Halobacterium halobium by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, the solubilization with 0,5 % (w/v SDS-Na and subsequent fractionation by methanol and gel filtration chromatography on Sephadex G-200 Column balanced with 0.09 M Tris-borate buffer (pH = 8,35 with 0,1 % (w/v SDS-Na and 2,5 mM EDTA. Within the framework of the research the mechanism of color perception by the visual analyzer having the ability to analyze certain ranges of the optical spectrum, as colors was studied along with an analysis of the additive mixing of two colors. It was shown that at the mixing of electromagnetic waves with different wavelengths, the visual analyzer perceive them as separate or average wave length corresponding to mix color.

  8. Calibration of Membrane Viscosity of the Reconstituted Vesicles by Measurement of the Rotational Diffusion of Bacteriorhodopsin

    Institute of Scientific and Technical Information of China (English)

    王敖金; 胡坤生

    2002-01-01

    Membrane viscosity of the reconstituted vesicles was calibrated by rotational diffusion of bacteriorhodopsin (BR) in dimyristoylphosphatidylcholine (DMPC) and egg phosphatidylcholine (PC) vesicles. Rotational diffusion of BR in the vesicles was measured by flash-induced absorption anisotropy decay. BR was, for the first time, reconstituted successfully into DMPC and egg PC vesicles. From the measurement of flash-induced absorption anisotropy decay of BR, the value of rotational diffusion coefficient D was obtained from each curve fitting by a global fitting procedure and, in turn, membrane viscosity η was estimated from D. The results have shown that membrane viscosity is temperature-dependent. It was decreased as temperature increased, but a transition occurred in the region of the respective phase transition of DMPC and egg PC, respectively. The decrease of η was fast near the phase transition for DMPC and egg PC. Few effects of lipid/BR ratio and glycerol or sucrose in suspension medium on membrane viscosity were found.

  9. Solid-state NMR studies of bacteriorhodopsin and the purple membrane

    CERN Document Server

    Mason, A J

    2001-01-01

    proteins. This technique may prove particularly useful when studying large proteins that are difficult to orient where the MAS lineshapes will remain relatively unaffected in comparison with current static NMR methods. Finally the MAOSS method was extended to the study of the lipid components of the purple membrane and the feasibility of determining structural constraints from phospholipid headgroups was assessed. The potential of using sup 3 sup 1 P NMR to observe qualitative protein-lipid interactions in both the purple membrane and reconstituted membranes containing bovine rhodopsin was also demonstrated. Following the demonstration of a new MAS NMR method for resolving orientational constraints in uni-axially oriented biological membranes (Glaubitz and Watts, 1998), experiments were performed to realise the potential of the new method on large, oriented membrane proteins. Using bacteriorhodopsin in the purple membrane as a paradigm for large membrane proteins, the protein was specifically labelled with de...

  10. Generation and analysis of bacteriorhodopsin mutants with the potential for biotechnological applications.

    Science.gov (United States)

    Saeedi, P; Moosaabadi, J Mohammadian; Sebtahmadi, S Sina; Mehrabadi, J Fallah; Behmanesh, M; Nejad, H Rouhani; Nazaktabar, A

    2012-01-01

    The properties of bacteriorhodopsin (BR) can be manipulated by genetic engineering. Therefore, by the methods of gene engineering, Asp85 was replaced individually by two other amino acids (D85V, D85S). The resulting recombinant proteins were assembled into soybean vesicles retinylated to form functional BR-like nano-particles. Proton translocation was almost completely abrogated by the mutant D85S, while the D85V mutant was partially active in pumping protons. Compared with wild type, maximum absorption of the mutants, D85V and D85S, were 563 and 609 nm, which illustrated 5 nm reductions (blue shift) and 41 nm increases (red shift), respectively. Since proton transport activity and spectroscopic activities of the mutants are different, a wide variety of membrane bioreactors (MBr) have been developed. Modified proteins can be utilized to produce unique photo/Electro-chromic materials and tools. PMID:22976247

  11. The effect of charged lipids on bacteriorhodopsin membrane reconstitution and its photochemical activities

    International Nuclear Information System (INIS)

    Bacteriorhodopsin (BR) was reconstituted into artificial lipid membrane containing various charged lipid compositions. The proton pumping activity of BR under flash and continuous illumination, proton permeability across membrane, as well as the decay kinetics of the photocycle intermediate M412 were studied. The results showed that lipid charges would significantly affect the orientation of BR inserted into lipid membranes. In liposomes containing anionic lipids, BRs were more likely to take natural orientation as in living cells. In neutral or positively charged liposomes, most BRs were reversely assembled, assuming an inside out orientation. Moreover, the lipids charges also affect BR's M intermediate kinetics, especially the slow component in M intermediate decay. The half-life M412s increased significantly in BRs in liposomes containing cationic lipids, while decreased in those in anionic liposomes

  12. Steady-State Characterization of Bacteriorhodopsin-D85N Photocycle

    Science.gov (United States)

    Timucin, Dogan A.; Downie, John D.; Norvig, Peter (Technical Monitor)

    1999-01-01

    An operational characterization of the photocycle of the genetic mutant D85N of bacteriorhodopsin, BR-D85N, is presented. Steady-state bleach spectra and pump-probe absorbance data are obtained with thick hydrated films containing BR-D85N embedded in a gelatin host. Simple two- and three-state models are used to analyze the photocycle dynamics and extract relevant information such as pure-state absorption spectra, photochemical-transition quantum efficiencies, and thermal lifetimes of dominant states appearing in the photocycle, the knowledge of which should aid in the analysis of optical recording and retrieval of data in films incorporating this photochromic material. The remarkable characteristics of this material and their implications from the viewpoint of optical data storage and processing are discussed.

  13. High-effective cultivation of Halobacterium salinarum providing with bacteriorhodopsin production under controlled stress.

    Science.gov (United States)

    Kalenov, Sergei V; Baurina, Marina M; Skladnev, Dmitry A; Kuznetsov, Alexander Ye

    2016-09-10

    Submerged growth of Halobacterium salinarum and therefore synthesis of bacteriorhodopsin (BR) and carotenoids depend greatly on products of both chemical and/or photochemical oxidation of medium components and cellular metabolism which act as inhibitors. Some cultivation variants which allowed eliminating an adverse effect of inhibitors on biomass accumulation and BR synthesis are reviewed. The application of activated charcoal or ion exchange resin as adsorbents at preparing inoculums and the main cultivation stages was shown to allow controlling, namely lowering overstress of the halobacterial cells by metabolites. The halobacterial biomass containing BR up to 1,750mgL(-1) and the minimum amount of carotinoids that would BR greatly facilitate isolation was accumulated up to 45gL(-1) during eight-day cultivation with cell recycling through adsorbent suspension in a fed-batch mode. To control BR biosynthesis the express method of BR quantification based on colour shades of cell suspension was developed. PMID:27449487

  14. High-speed integrated optical logic based on the protein bacteriorhodopsin.

    Science.gov (United States)

    Mathesz, Anna; Fábián, László; Valkai, Sándor; Alexandre, Daniel; Marques, Paulo V S; Ormos, Pál; Wolff, Elmar K; Dér, András

    2013-08-15

    The principle of all-optical logical operations utilizing the unique nonlinear optical properties of a protein was demonstrated by a logic gate constructed from an integrated optical Mach-Zehnder interferometer as a passive structure, covered by a bacteriorhodopsin (bR) adlayer as the active element. Logical operations were based on a reversible change of the refractive index of the bR adlayer over one or both arms of the interferometer. Depending on the operating point of the interferometer, we demonstrated binary and ternary logical modes of operation. Using an ultrafast transition of the bR photocycle (BR-K), we achieved high-speed (nanosecond) logical switching. This is the fastest operation of a protein-based integrated optical logic gate that has been demonstrated so far. The results are expected to have important implications for finding novel, alternative solutions in all-optical data processing research. PMID:23500476

  15. Broadband optical limiter based on nonlinear photoinduced anisotropy in bacteriorhodopsin film

    Science.gov (United States)

    Huang, Yuhua; Siganakis, Georgios; Moharam, M. G.; Wu, Shin-Tson

    2004-11-01

    Nonlinear photoinduced anisotropy in a bacteriorhodopsin film was theoretically and experimentally investigated and a broadband active optical limiter was demonstrated in the visible spectral range. A diode-pumped second harmonic yttrium aluminum garnet laser was used as a pumping beam and three different wavelengths at λ =442, 532, and 655nm from different lasers were used as probing beams. The pump and probe beams overlap at the sample. When the pumping beam is absent, the probing beam cannot transmit the crossed polarizers. With the presence of the pumping beam, a portion of the probing light is detected owing to the photoinduced anisotropy. Due to the optical nonlinearity, the transmitted probing beam intensity is clamped at a certain value, which depends on the wavelength, when the pumping beam intensity exceeds 5mW/mm2. Good agreement between theory and experiment is found.

  16. An all-optical time-delay relay based n a bacteriorhodopsin film

    Institute of Scientific and Technical Information of China (English)

    Chen Gui-Ying; Xu Xu-Xu; Zhang Chun-Ping; Qi Shen-Wen; Song Qi-Wang

    2008-01-01

    Using a special property of dynamic complementary-suppression-modulated transmission (DCSMT) in the bacteriorhodopsin (bR) film,we have demonstrated an all-optical time-delay relay.To extend our work,the relationship between the delay time of the all-optical time-delay relay and parameters of a bR film is numerically studied.We show how the delay time changes with the product of concentration and thickness (PCT) of a bR film.Furthermore,the shortest and longest delay times are given for the relay of 'switch off'.The saturable delay time and maximum delaytime of 'switch on' are also given.How the wavelengths (632.8,568,533 and 412 nm) and intensities of the illuminating light influence the delay time is also discussed.The simulation results are useful for optimizing the design of all-optical time-delay relays.

  17. All-optical switching characteristics in bacteriorhodopsin and its applications in integrated optics

    Science.gov (United States)

    Huang, Yuhua; Wu, Shin-Tson; Zhao, Youyuan

    2004-03-01

    We experimentally and theoretically investigated the optical switching characteristics of bacteriorhodopsin (bR) at l=633 nm using the pump-probe method. A diode-pumped second harmonic YAG laser (l=532 nm which is located around the maximum initial Br state absorption) was used as a pumping beam and a cw He-Ne laser (l=633 nm which is around the peaks of K and O states) was used as a probe. Due to the nonlinear intensity induced excited state absorption of the K, L, M, N, and O states in the bR photocycle, the switching characteristics are sensitive to the intensity of the probe and pump beams. Based on this property, we have demonstrated an all-optical device functioning as 11 kinds of variable binary all-optical logic gates.

  18. Photoinduced nonlinear refraction in a polymeric film encapsulating a bacteriorhodopsin mutant

    Science.gov (United States)

    Teng, Xue-lei; Lu, Ming; Zhao, You-yuan; Ma, De-wang; Zhao, Ying-chun; Ding, Jian-dong; Huang, Wei-da

    2010-08-01

    The absorption change versus time after switching off the illumination on a polymeric film that contained a bacteriorhodopsin (BR) mutant has been measured. The M-lifetime of this BR mutant is ˜320 s. A pertinent Z-scan was performed to study the BR optical nonlinearity. A physical model with multi-level transitions in the Z-scan was suggested. The minimum saturated light intensity measured at 633 nm is ˜0.9 mW/cm2. A low intensity of 70 μW/cm2 has been used for recording in this film. The change in refraction index Δn633 is -3.0×10-3 and Δn476 is 8.5×10-3 with the intensity all at ˜100 mW/cm2.

  19. Cooperative phenomena in the photocycle of D96N mutant bacteriorhodopsin.

    Science.gov (United States)

    Radionov, A N; Kaulen, A D

    1995-12-27

    The M intermediate decay in the photocycle of D96N mutant bacteriorhodopsin does not depend on the light intensity of the exciting flash. Cooperative phenomena in the photocycle are revealed after addition of azide causing acceleration of the M decay and making it kinetically well separated from the N decay. Increase in the light intensity induces slight deceleration of the M decay and significant acceleration of the N decay. The data obtained directly confirm our recent model [Komrakov and Kaulen (1995) Biophys. Chem. 56, 113-119], according to which appearance of the Mslow intermediate in the photocycle of the wild type bR at high light intensity is due to destabilization of the N intermediate leading to the acceleration of the N-->M and N-->bR reactions. PMID:8549749

  20. Systems Issues Pertaining to Holographic Optical Data Storage in Thick Bacteriorhodopsin Films

    Science.gov (United States)

    Downie, John D.; Timucin, Dogan A.; Gary, Charles K.; Oezcan, Meric; Smithey, Daniel T.; Crew, Marshall; Lau, Sonie (Technical Monitor)

    1998-01-01

    The optical data storage capacity and raw bit-error-rate achievable with thick photochromic bacteriorhodopsin (BR) films are investigated for sequential recording and read- out of angularly- and shift-multiplexed digital holograms inside a thick blue-membrane D85N BR film. We address the determination of an exposure schedule that produces equal diffraction efficiencies among each of the multiplexed holograms. This exposure schedule is determined by numerical simulations of the holographic recording process within the BR material, and maximizes the total grating strength. We also experimentally measure the shift selectivity and compare the results to theoretical predictions. Finally, we evaluate the bit-error-rate of a single hologram, and of multiple holograms stored within the film.

  1. On the mechanism of weak-field coherent control of retinal isomerization in bacteriorhodopsin

    International Nuclear Information System (INIS)

    Experimental studies of the short time reaction dynamics controlling the chemical branching ratio provide direct evidence for the mechanism of coherent control of the retinal photoisomerization in bacteriorhodopsin in the weak-field limit with respect to the previous report [V. Prokhorenko, A. Nagy, S. Waschuk, L. Brown, R. Birge, R. Miller, Science 313 (2006) 1257]. The phase sensitivity of the reaction dynamics is directly revealed using time- and frequency-resolved pump-probe measurements. The high degree of control of the reaction branching ratio is theoretically explained through a combination of spectral amplitude shaping and phase-dependent coupling to selectively excite vibrations most strongly coupled to the reaction coordinate. Coherent control in this context must involve reaction dynamics that occur on time scales comparable to electronic and vibrational decoherence time scales

  2. Effective atomic numbers and electron densities of bacteriorhodopsin and its comprising amino acids in the energy range 1 keV–100 GeV

    Energy Technology Data Exchange (ETDEWEB)

    Ahmadi, Morteza; Lunscher, Nolan [Waterloo Institute for Nanotechnology and Department of Systems Design Engineering, University of Waterloo, 200 University Ave., W., Waterloo, Ontario, Canada N2L 3G1 (Canada); Yeow, John T.W., E-mail: jyeow@uwaterloo.ca [Waterloo Institute for Nanotechnology and Department of Systems Design Engineering, University of Waterloo, 200 University Ave., W., Waterloo, Ontario, Canada N2L 3G1 (Canada)

    2013-04-01

    Recently, there has been an interest in fabrication of X-ray sensors based on bacteriorhodopsin, a proton pump protein in cell membrane of Halobacterium salinarium. Therefore, a better understanding of interaction of X-ray photons with bacteriorhodopsin is required. We use WinXCom program to calculate the mass attenuation coefficient of bacteriorhodopsin and its comprising amino acids for photon energies from 1 keV to 100 GeV. These amino acids include alanine, arginine, asparagine, aspartic acid, glutamine, glutamic acid, glycine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, Asx1, Asx2, Glx1 and Glx2. We then use that data to calculate effective atomic number and electron densities for the same range of energy. We also emphasize on two ranges of energies (10–200 keV and 1–20 MeV) in which X-ray imaging and radiotherapy machines work.

  3. Enhanced photocurrent generation in bacteriorhodopsin based bio-sensitized solar cells using gel electrolyte.

    Science.gov (United States)

    Chellamuthu, Jeganathan; Nagaraj, Pavithra; Chidambaram, Sabari Girisun; Sambandam, Anandan; Muthupandian, Ashokkumar

    2016-09-01

    High purity light sensitive photoactive protein Bacteriorhodopsin (BR) was isolated successfully via a simple two phase extraction technique (ATPS) as an alternate method for the tedious sucrose gradient ultracentrifugation procedure (SGU). Bio sensitized solar cells (BSSCs) were fabricated by the integration of BR into TiO2 (photo anode) with acetamide based gel electrolytes and platinum (photo cathode) as a counter electrode. The structural and photoelectrical behaviours of BR and BSSCs were analyzed by Atomic Force Microscopy, Raman spectroscopy, photocurrent and photovoltage (IV) measurement and electrochemical impedance spectroscopy. The short circuit photocurrent (Jsc) and photoelectric conversion efficiency (η) of acetamide based gel electrolyte (AG) (1.08mAcm(-2), 0.49%) are twice higher than that of traditional triiodide based liquid electrolyte (LE) (0.62mAcm(-2), 0.19%). Also, quasi-Fermi level and lifetime of photogenerated electrons in acetamide based gel electrolyte is about four times higher than that observed in traditional triiodide redox electrolyte. A comparison of the observed results with similar BSSCs made of other natural photoactive protein systems shows that BR as sensitizer has better photovoltaic performance. The enhanced photocurrent generation of the BSSC constructed in our study could be due to the interaction of BR with acetamide based modified poly(ethylene)oxide (PEO) gel electrolyte. PMID:27380296

  4. Schiff base switch II precedes the retinal thermal isomerization in the photocycle of bacteriorhodopsin.

    Directory of Open Access Journals (Sweden)

    Ting Wang

    Full Text Available In bacteriorhodopsin, the order of molecular events that control the cytoplasmic or extracellular accessibility of the Schiff bases (SB are not well understood. We use molecular dynamics simulations to study a process involved in the second accessibility switch of SB that occurs after its reprotonation in the N intermediate of the photocycle. We find that once protonated, the SB C15 = NZ bond switches from a cytoplasmic facing (13-cis, 15-anti configuration to an extracellular facing (13-cis, 15-syn configuration on the pico to nanosecond timescale. Significantly, rotation about the retinal's C13 = C14 double bond is not observed. The dynamics of the isomeric state transitions of the protonated SB are strongly influenced by the surrounding charges and dielectric effects of other buried ions, particularly D96 and D212. Our simulations indicate that the thermal isomerization of retinal from 13-cis back to all-trans likely occurs independently from and after the SB C15 = NZ rotation in the N-to-O transition.

  5. Surface modification by using of immobilized electrostatic self-assembly of bacteriorhodopsin as protein memory

    Directory of Open Access Journals (Sweden)

    Ashkan Zare Karizak

    2014-11-01

    Full Text Available Bacteriorhodopsin (BR is the light harvesting and photoactive proton pump found in the membrane of a salt marsh bacteria. This protein has significant potential to use in optical computing and memory devices due to unique intrinsic physical properties of photo and bioelectric. All these features make BR one of the most promising protein candidates in protein memories. Protein memory is a kind of optical memory with a large storage capacity and high speed processing features. BR protein was used with the polymer film in order to create better stability. In order to investigate immobilization of electrostatic self-assembly of BR on glass and polycarbonate as protein memories was used. Polycarbonate is a layer of compact disc (CD structure which considered dye immobilized on its surface and have reading and writing abilities of information via 0,1 bites. In this study, surfaces of polycarbonate modified by the mixture of 5% sulfuric acid and 20% acetic acid; furthermore, by using of PEI as cationic resin the surface of polycarbonate was charged and BR immobilized on it electrostatically. The modified surfaces were characterized by AFM technique. Also, light activity for reading data is retained. This is an appropriate method for optimal stability and activity assay of the protein and also is suitable for preparation of protein memories.

  6. Bacteriorhodopsin-based Langmuir-Schaefer films for solar energy capture.

    Science.gov (United States)

    Bertoncello, Paolo; Nicolini, Davide; Paternolli, Cristina; Bavastrello, Valter; Nicolini, Claudio

    2003-06-01

    The photovoltaic (PV) solar cell, converting incident solar radiation directly into electrical energy, today represents the most common power source for the earth-orbiting spacecraft, and the utilization of organic materials in this context is here explored in comparison with the present state of the art placing emphasis in organic nanotechnology. Poly[3-3'(vinylcarbazole)] (PVK) was synthesized by oxidative polymerization with ferric chloride of N-vinylcarbazole. The resulting polymer was then deposited on solid support by using the Langmuir-Schaefer (LS) technique. The pressure-area isotherm of PVK revealed the possibility of compact monolayer formation at the air-water interface. Different layers of PVK were doped with iodine vapors. The cyclic voltammetry investigation of PVK-doped I2 showed a distinctive electrochemical behavior. The photoinduced charge transfer across a donor/acceptor (D/A) hybrid interface provided an effective method to study the PV properties of the composite LS films. The results are compared with other approaches within the biological framework, such as bacteriorhodopsin (BR), and organic nanostructured materials. PMID:15382669

  7. Kinetics of picosecond laser pulse induced charge separation and proton transfer in bacteriorhodopsin.

    Science.gov (United States)

    Yao, Baoli; Xu, Dalun; Hou, Xun; Hu, Kunsheng; Wang, Aojin

    2003-01-01

    Bacteriorhodopsin (BR) films oriented by an electrophoretic method are deposited on a transparent conductive ITO glass. A counterelectrode of copper and gelose gel is used to compose a sandwich-type photodetector with the structure of ITO/BR film/gelose gel/Cu. A single 30-ps laser pulse and a mode-locked pulse train are respectively used to excite the BR photodetector. The ultrafast falling edge and the bipolar response signal are measured by the digital oscilloscope under seven different time ranges. Marquardt nonlinear least squares fitting is used to fit all the experimental data and a good fitting equation is found to describe the kinetic process of the photoelectric signal. Data fitting resolves six exponential components that can be assigned to a seven-step BR photocycle model: BR-->K-->KL-->L-->M-->N-->O-->BR. Comparing tests of the BR photodetector with a 100-ps Si PIN photodiode demonstrates that this type of BR photodetector has at least 100-ps response time and can also serve as a fast photoelectric switch. PMID:12542379

  8. B-M-type anisotropy in bacteriorhodopsin films for nonlinear spatial light modulation

    Science.gov (United States)

    Korchemskaya, Elena Y.; Stepanchikov, Dmitriy A.; Dyukova, Tatyana V.; Shakhbazian, Valery Y.

    2003-02-01

    Bacteriorhodopsin (BR), a photoreceptor protein possesses a photochemical cycle of several distinct intermediates; all of them are photoactive. The BR molecules both in the initial form of the photocycle, BR570 (absorption maximum around 570 nm) and longest-lived (in films) intermediate M412 (absorption maximum at 412 nm) possess anisotropic absorption. Under the action of linearly polarized light, the reversible anisotropic photoselection of BR molecules takes place. So far only the method of photoinduced anisotropy based on anisotropic properties of BR570 was applied to realtime optical processing. In the present work, the potentialities for the use of photoinduced anisotropy in the BR-films based on both BR570 and M412 for the spatial light modulation are demonstrated. The overall blocking of highintensity features from an image is shown. Mixed B-M-type anisotropy in the chemically modified BR films, as applied to the edge enhancement, can provide a contrast ratio as high as 250:1. Low saturation intensity of the BR-films allows for the blocking of any intensity feature from an image that is carried out by choosing an appropriate intensity level of a controlling He-Ne laser beam without analyzer rotation. The photoanisotropic incoherent-to-coherent optical conversion with concurrent spatial-intensity modulation is also performed on the BR-films.

  9. Polarization multiplexed write-once-read-many optical data storage in bacteriorhodopsin films

    Science.gov (United States)

    Yao, Baoli; Lei, Ming; Ren, Liyong; Menke, Neimule; Wang, Yingli; Fischer, Thorsten; Hampp, Norbert

    2005-11-01

    In polymeric films of bacteriorhodopsin (BR) a photoconversion product, which was named the F620 state, was observed on excitation of the film with 532 nm nanosecond laser pulses. This photoproduct shows a strong nonlinear absorption. Such BR films can be used for write-once-read-many (WORM) optical data storage. We demonstrate that a photoproduct similar or even identical to that obtained with nanosecond pulses is generated on excitation with 532 nm femtosecond pulses. This photoproduct also shows strong anisotropic absorption, which facilitates polarization storage of data. The product is thermally stable and is irretrievable to the initial B state either by photochemical reaction or through a thermal pathway. The experimental results indicate that the product is formed by a two-photon absorption process. Optical WORM storage is demonstrated by use of two polarization states, but more polarization states may be used. The combination of polarization data multiplexing and extremely short recording time in the femtosecond range enables very high data volumes to be stored within a very short time.

  10. All-optical switching and all-optical logic gates based on bacteriorhodopsin

    Science.gov (United States)

    Huang, Yuhua; Wu, Shin-Tson; Zhao, Youyuan

    2004-06-01

    We demonstrate an all-optical switching using a bacteriorhodopsin (bR) film. The transmission of the bR film is investigated using the pump-probe method. A diode-pumped second harmonic YAG laser (λ = 532nm which is around the maximum initial B state absorption) was used as a pumping beam and a cw He-Ne laser (λ = 632 nm which is around the peaks of K and O states) was used as a probe. Due to the nonlinear intensity induced excited state absorption of the K, L, M, N, and O states in the bR photocycle, the switching characteristics are sensitive to the intensity of the probe and pump beams. Based on this property, we design an all-optical operating device functioning as 11 kinds of variable binary all-optical logic gates. The incident 532nm beam acts as an input to the logic gate and the transmission of the 632nm bears the output of the gate.

  11. Photosensory behaviour of a bacteriorhodopsin-deficient mutant, ET-15, of Halobacterium halobium

    International Nuclear Information System (INIS)

    Halobacterium halobium, strain ET-15, which does not contain detectable amounts of bacteriorhodopsin (BR) shows behavioral responses to UV and yellow-green light. Attractant stimuli, i.e. light-increases in the yellow-green range or light-decreases in the UV, suppress the spontaneous reversals of the swimming direction for a certain time. Repellent stimuli, i.e. light-decreases in the yellow-green range or light-increases in the UV, elicit an additional reversal response after a few seconds. Action spectra of both sensory photosystems, PS 370 and PS 565, were measured with attractant as well as with repellent stimuli. As in BR-containing cells, maximal sensitivity was always found at 370 nm for the UV-system and at 565 nm for the long-wavelength system. Fluence-response curves at 370 and 565 nm obtained with strain ET-15 and with a BR-containing strain show that the sensitivity of both photosystems is not reduced in the absence of BR. It is concluded that BR is required neither for PS 565 nor for PS 370. Instead retinal-containing pigments different from BR have to be assumed to mediate photosensory behavior. (author)

  12. Förster Resonance Energy Transfer between Core/Shell Quantum Dots and Bacteriorhodopsin

    Directory of Open Access Journals (Sweden)

    Mark H. Griep

    2012-01-01

    Full Text Available An energy transfer relationship between core-shell CdSe/ZnS quantum dots (QDs and the optical protein bacteriorhodopsin (bR is shown, demonstrating a distance-dependent energy transfer with 88.2% and 51.1% of the QD energy being transferred to the bR monomer at separation distances of 3.5 nm and 8.5 nm, respectively. Fluorescence lifetime measurements isolate nonradiative energy transfer, other than optical absorptive mechanisms, with the effective QD excited state lifetime reducing from 18.0 ns to 13.3 ns with bR integration, demonstrating the Förster resonance energy transfer contributes to 26.1% of the transferred QD energy at the 3.5 nm separation distance. The established direct energy transfer mechanism holds the potential to enhance the bR spectral range and sensitivity of energies that the protein can utilize, increasing its subsequent photocurrent generation, a significant potential expansion of the applicability of bR in solar cell, biosensing, biocomputing, optoelectronic, and imaging technologies.

  13. Deposition of Bacteriorhodopsin Protein in a Purple Membrane Form on Nitrocellulose Membranes for Enhanced Photoelectric Response

    Science.gov (United States)

    Kim, Young Jun; Neuzil, Pavel; Nam, Chang-Hoon; Engelhard, Martin

    2013-01-01

    Bacteriorhodopsin protein (bR)-based systems are one of the simplest known biological energy converters. The robust chemical, thermal and electrochemical properties of bR have made it an attractive material for photoelectric devices. This study demonstrates the photoelectric response of a dry bR layer deposited on a nitrocellulose membrane with indium tin oxide (ITO) electrodes. Light-induced electrical current as well as potential and impedance changes of dried bR film were recorded as the function of illumination. We have also tested bR in solution and found that the electrical properties are strongly dependent on light intensity changing locally proton concentration and thus pH of the solution. Experimental data support the assumption that bR protein on a positively charged nitrocellulose membrane (PNM) can be used as highly sensitive photo- and pH detector. Here the bR layer facilitates proton translocation and acts as an ultrafast optoelectric signal transducer. It is therefore useful in applications related to bioelectronics, biosensors, bio-optics devices and current carrying junction devices. PMID:23271605

  14. Enhanced Photocurrent Generation from Bacteriorhodopsin Photocells Using Grating-Structured Transparent Conductive Oxide Electrodes.

    Science.gov (United States)

    Kaji, Takahiro; Kasai, Katsuyuki; Haruyama, Yoshihiro; Yamada, Toshiki; Inoue, Shin-Ichiro; Tominari, Yukihiro; Ueda, Rieko; Terui, Toshifumi; Tanaka, Shukichi; Otomo, Akira

    2016-04-01

    We fabricated a grating-structured electrode made of indium-doped zinc oxide (IZO) with a high refractive index (approximately 2) for a bacteriorhodopsin (bR) photocell. We investigated the photocurrent characteristics of the bR photocell and demonstrated that the photocurrent values from the bR/IZO electrode with the grating structure with a grating period of 340 nm were more than 3.5-4 times larger than those without the grating structure. The photocurrent enhancement was attributed to the resonance effect due to light coupling to the grating structure as well as the scattering effect based on the experimental results and analysis using the photonic band structure determined using finite-difference time-domain (FDTD) simulations. The refractive index of the bR film in electrolyte solution (1.40) used in the FDTD simulations was estimated by analyzing the extinction peak wavelength of 20-nm gold colloids in the bR film. Our results indicate that the grating- or photonic-crystal-structured transparent conductive oxide (TCO) electrodes can increase the light use efficiency of various bR devices such as artificial photosynthetic devices, solar cells, and light-sensing devices. PMID:27451605

  15. Application of chemical modification and spin-labeling techniques to the study of energy conversion by bacteriorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Packer, L.; Quintanilha, A.T.; Mehlhorn, R.J.

    1983-03-01

    Light generates a pH gradient and an electrical potential across the purple membrane of Halobacterium halobium. We are investigating the time-resolved changes in protonation of the side chains of specific amino-acid residues and the correlation of these changes with photon absorption and the ensuing photo-reaction cycle. We seek to determine the precise molecular description of the photocycle and of the time dependent steps in the uptake, translocation, and release of protons by the retinal proton catalyst in this membrane, bacteriorhodopsin (BR). 14 references, 3 figures, 1 table.

  16. Nonlinear optical method for the investigation of spectral properties of biomolecular complexes: second harmonic generation in ordered structures of bacteriorhodopsin

    Science.gov (United States)

    Aktsipetrov, Oleg A.; Fedyanin, Andrew A.; Murzina, Tatyana V.; Borisevich, G. P.; Kononenko, A. A.

    1995-02-01

    For the first time the method of the second harmonic generation was used to study the photo- and electrically induced nonlinear optical transformations in thin oriented films of purple membranes (PM). Variations of the film nonlinear susceptibility were investigated as the bacteriorhodopsin (bR) molecule underwent the cycle of photoinduced transformations for both dry electrically oriented films and bR molecules embedded into poly(vinyl alcohol) matrix. The electrically induced changes of the nonlinear optical properties were studied for the electrostatic field strength up to the values 4 (DOT) 104 V/cm. Nonlinear susceptibilities of oriented and nonoriented dried PM films are compared.

  17. Application of nonlinear absorption properties and light adaptation process in the polymer films based on bacteriorhodopsin for the low-power optical signal processing

    Science.gov (United States)

    Korchemskaya, Elena Y.; Soskin, Marat S.

    1994-01-01

    Experimental and theoretical investigations were made of the characteristics of nonlinear transmission and light adaptation processes of polymer films based on bacteriorhodopsin (BR). It was found that media containing BR can be used to enhance the contrast of low-power signals for realization of the connection structure of the neural network.

  18. High resolution electron diffraction analysis of structural changes associated with the photocycle of bacteriorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Han, B. -G. [Lawrence Berkeley Lab., CA (United States). Life Sciences Div.; Univ. of California, Berkeley, CA (United States). Dept. of Biophysics

    1994-04-01

    Changes in protein structure that occur during the formation of the M photointermediate of bacteriorhodopsin can be directly visualized by electron diffraction techniques. Samples containing a high percentage of the M intermediate were trapped by rapidly cooling the crystals with liquid nitrogen following illumination with filtered green light at 240K and 260K respectively. Difference Fourier projection maps for M minus bR at two temperatures and for M{sub 260K} minus M{sub 240K} are presented. While it is likely that a unique M-substate is trapped when illuminated at 260K produces a mixture of the M{sub 240K} substate and a second M-substate which may have a protein structure similar to the N-intermediate. The diffraction data clearly show that statistically significant structural changes occur upon formation of the M{sub 240K} specimen and then further upon formation of the second substate which is present in the mixture that is produced at 260K. A preliminary 3-D difference map, based on data collected with samples tilted up to 30{degree}, has been constructed at a resolution of 3.5{angstrom} parallel to the membrane plane and a resolution of 8.5{angstrom} perpendicular to the membrane. The data have been analyzed by a number of different criteria to ensure that the differences seen reflect real conformation changes at a level which is significantly above the noise in the map. Furthermore, a comparison of the positions of specific backbone and side-chain groups relative to significant difference peaks suggests that it will be necessary to further refine the atomic resolution model before it will be possible to interpret the changes in chemical structure that occur in the protein at this stage of the photocycle.

  19. Coherent control of the isomerization of retinal in bacteriorhodopsin in the high intensity regime

    International Nuclear Information System (INIS)

    Coherent control protocols provide a direct experimental determination of the relative importance of quantum interference or phase relationships of coupled states along a selected pathway. These effects are most readily observed in the high intensity regime where the field amplitude is sufficient to overcome decoherence effects. The coherent response of retinal photoisomerization in bacteriorhodopsin to the phase of the photoexcitation pulses was examined at fluences of 1015- 2.5 x 1016 photons per square centimeter, comparable to or higher than the saturation excitation level of the S0-S1 retinal electronic transition. At moderate excitation levels of ∼6 x 1015 photons/cm2 (2), chirping the excitation pulses increases the all-trans to 13-cis isomerization yield by up to 16% relative to transform limited pulses. The reported results extend previous weak-field studies [Prokhorenko et al., Science 313, 1257 (2006)] and further illustrate that quantum coherence effects persist along the reaction coordinate in strong fields even for systems as complex as biological molecules. However, for higher excitation levels of ∼200 GW/cm2, there is a dramatic change in photophysics that leads to multiphoton generated photoproducts unrelated to the target isomerization reaction channel and drastically changes the observed isomerization kinetics that appears, in particular, as a red shift of the transient spectra. These results explain the apparent contradictions of the work by Florean et al.[Proc. Natl. Acad. Sci. U.S.A. 106, 10896 (2009)] in the high intensity regime. We are able to show that the difference in observations and interpretation is due to artifacts associated with additional multiphoton-induced photoproducts. At the proper monitoring wavelengths, coherent control in the high intensity regime is clearly observable. The present work highlights the importance of conducting coherent control experiments in the low intensity regime to access information on quantum

  20. Optical Fourier and Holographic Techniques for Medical Image Processing with Bacteriorhodopsin

    Science.gov (United States)

    Yelleswarapu, Chandra

    2008-03-01

    The biological photochrome bacteriorhodopsin (bR) shows many intrinsic optical and physical properties. The active chromophore in bR is a retinal group which absorbs light and goes through a photocycle. The unique feature of the system is its flexibility -- the photocycle can be optically controllable since the process of photoisomerization can go in both directions depending on wavelength, intensity and polarization of the incident light, opening a variety of possibilities for manipulating amplitude, phase, polarization and index of refraction of the incident light. Over the years we studied the basic nonlinear optics and successfully exploited the unique properties for several optical spatial filtering techniques with applications in medical image processing. For nonlinear Fourier filtering, the photo-controlled light modulating characteristics of bR films are exploited. At the Fourier plane, the spatial frequency information carried by a blue probe beam at 442 nm is selectively manipulated in the bR film by changing the position and intensity of a yellow control beam at 568 nm. In transient Fourier holography, photoisomerizative gratings are recorded and reconstructed in bR films. Desired spatial frequencies are obtained by matching the reference beam intensity to that of the particular frequency band in object beam. A novel feature of the technique is the ability to transient display of selected spatial frequencies in the reconstructing process which enables radiologists to study the features of interest in time scale. The results offer useful information to radiologists for early detection of breast cancer. Some of the highlights will be presented.

  1. All-Optical Switching in Bacteriorhodopsin Based on Excited-State Absorption

    Science.gov (United States)

    Roy, Sukhdev

    2008-03-01

    Switching light with light is of tremendous importance for both fundamental and applied science. The advent of nano-bio-photonics has led to the design, synthesis and characterization of novel biomolecules that exhibit an efficient nonlinear optical response, which can be utilized for designing all-optical biomolecular switches. Bacteriorhodopsin (bR) protein found in the purple membrane of Halobacterium halobium has been the focus of intense research due to its unique properties that can also be tailored by physical, chemical and genetic engineering techniques to suit desired applications. The talk would focus on our recent results on all-optical switching in bR and its mutants, based on excited-state absorption, using the pump-probe technique. We would discuss the all-optical control of various features of the switching characteristics such as switching contrast, switching time, switching pump intensity, switched probe profile and phase, and relative phase-shift. Optimized conditions for all-optical switching that include optimized values of the small-signal absorption coefficient (for cw case), the pump pulse width and concentration for maximum switching contrast (for pulsed case), would be presented. We would discuss the desired optimal spectral and kinetic properties for device applications. We would also discuss the application of all-optical switching to design low power all-optical computing devices, such as, spatial light modulators, logic gates and multiplexers and compare their performance with other natural photoreceptors such as pharaonis phoborhodopsin, proteorhodopsin, photoactive yellow protein and the blue light plant photoreceptor phototropin.

  2. Studying the Mechanism of Phototransformation of Light Signal by Various Mammal and Bacterial Photoreceptor Pigments  Rhodopsin, Iodopsin and Bacteriorhodopsin

    Directory of Open Access Journals (Sweden)

    Ignat Ignatov

    2015-06-01

    Full Text Available This review article outlines the structure and function of mammal and bacterial photoreceptor pigments (rhodopsin, iodopsin, bacteriorhodopsin and their aspects of bio-nanotechnological usage. On an example of bacteriorhodopsin is described the method of its isolation from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, the solubilization with 0,5 % (w/v SDS-Na and subsequent fractionation by methanol and gel filtration chromatography on Sephadex G-200 Column balanced with 0,09 M Tris-buffer (pH = 8,35 with 0,1 % (w/v SDS-Na and 2,5 mM EDTA. Within the framework of the research the mechanism of color perception by the visual retina analyzer having the ability to analyze certain ranges of the optical spectrum as colors, was studied along with an analysis of the additive mixing of two or more colors. It was shown that at the mixing of electromagnetic waves with different wavelengths, the visual analyzer perceives them as the separate or average wave length corresponding to the mixing color.

  3. A residue substitution near the beta-ionone ring of the retinal affects the M substates of bacteriorhodopsin

    Science.gov (United States)

    Varo, G.; Zimanyi, L.; Chang, M.; Ni, B.; Needleman, R.; Lanyi, J. K.

    1992-01-01

    The switch in the bacteriorhodopsin photocycle, which reorients access of the retinal Schiff base from the extracellular to the cytoplasmic side, was suggested to be an M1----M2 reaction (Varo and Lanyi. 1991. Biochemistry. 30:5008-5015, 5016-5022). Thus, in this light-driven proton pump it is the interconversion of proposed M substates that gives direction to the transport. We find that in monomeric, although not purple membrane-lattice immobilized, D115N bacteriorhodopsin, the absorption maximum of M changes during the photocycle: in the time domain between its rise and decay it shifts 15 nm to the blue relative to the spectrum at earlier times. This large shift strongly supports the existence of two M substates. Since D115 is located near the beta-ionone ring of the retinal, the result raises questions about the possible involvement of the retinal chain or protein residues as far away as 10 A from the Schiff base in the mechanism of the switching reaction.

  4. Observation of helix associations for insertion of a retinal molecule and distortions of helix structures in bacteriorhodopsin

    Science.gov (United States)

    Urano, Ryo; Okamoto, Yuko

    2015-12-01

    We applied a newly proposed prediction method for membrane protein structures to bacteriorhodopsin that has distorted transmembrane helices in the native structure. This method uses an implicit membrane model, which restricts sampling space during folding in a membrane region, and includes helix bending. Replica-exchange simulations were performed with seven transmembrane helices only without a retinal molecule. Obtained structures were classified into clusters of similar structures, which correspond to local-minimum free energy states. The two lowest free energy states corresponded to a native-like structure with the correct empty space for retinal and a structure with this empty space filled with a helix. Previous experiments of bacteriorhodopsin suggested that association of transmembrane helices enables them to make a room for insertion of a retinal. Our results are consistent with these results. Moreover, distortions of helices in the native-like structures were successfully reproduced. In the distortions, whereas the locations of kinks for all helices were similar to those of Protein Data Bank's data, the amount of bends was more similar for helices away from the retinal than for those close to the retinal in the native structure. This suggests a hypothesis that the amino-acid sequence specifies the location of kinks in transmembrane helices and that the amount of distortions depends on the interactions with the surrounding molecules such as neighboring helices, lipids, and retinal.

  5. Proton uptake mechanism of bacteriorhodopsin as determined by time-resolved stroboscopic-FTIR-spectroscopy.

    Science.gov (United States)

    Souvignier, G; Gerwert, K

    1992-11-01

    Bacteriorhodopsin's proton uptake reaction mechanism in the M to BR reaction pathway was investigated by time-resolved FTIR spectroscopy under physiological conditions (293 K, pH 6.5, 1 M KCl). The time resolution of a conventional fast-scan FTIR spectrometer was improved from 10 ms to 100 mus, using the stroboscopic FTIR technique. Simultaneously, absorbance changes at 11 wavelengths in the visible between 410 and 680 nm were recorded. Global fit analysis with sums of exponentials of both the infrared and visible absorbance changes yields four apparent rate constants, k(7) = 0.3 ms, k(4) = 2.3 ms, k(3) = 6.9 ms, k(6) = 30 ms, for the M to BR reaction pathway. Although the rise of the N and O intermediates is dominated by the same apparent rate constant (k(4)), protein reactions can be attributed to either the N or the O intermediate by comparison of data sets taken at 273 and 293 K. Conceptionally, the Schiff base has to be oriented in its deprotonated state from the proton donor (asp 85) to the proton acceptor (asp 96) in the M(1) to M(2) transition. However, experimentally two different M intermediates are not resolved, and M(2) and N are merged. From the results the following conclusions are drawn: (a) the main structural change of the protein backbone, indicated by amide I, amide II difference bands, takes place in the M to N (conceptionally M(2)) transition. This reaction is proposed to be involved in the "reset switch" of the pump, (b) In the M to N (conceptionally M(2)) transition, most likely, asp-85's carbonyl frequency shifts from 1,762 to 1,753 cm(-1) and persists in O. Protonation of asp-85 explains the red-shift of the absorbance maximum in O. (c) The catalytic proton uptake binding site asp-96 is deprotonated in the M to N transition and is reprotonated in O. PMID:19431858

  6. Synthesis of 13C and 2H labelled retinals: spectroscopic investigations on isotopically labelled rhodopsin and bacteriorhodopsin

    International Nuclear Information System (INIS)

    In order to develop probes of the structure of chromophores, the author introduces isotopic modifications at specific chromophoric positions as structural probes. To obtain bacteriorhodopsin, rhodopsin and their photoproducts labelled in the chromophore at selected positions, bacterioopsin and opsin were reacted with the appropriate labelled a11-trans and 11-cis retinals. The author describes the synthesis of a11-trans retinal selectively 13C labelled at different positions. The characterization of these labelled a11-trans retinals by mass spectrometry, 300 MHz 1H NMR and 75 MHz 13C NMR spectroscopy is given. The photochemical preparation and isolation of the pure 9-, 11- and 13-cis forms is described in the experimental part. (Auth.)

  7. Towards structural investigations on isotope labelled native bacteriorhodopsin in detergent micelles by solution-state NMR spectroscopy

    International Nuclear Information System (INIS)

    1H NMR signals of the retinal moiety in detergent-solubilized bacteriorhodopsin are assigned, enabling the interpretation of NOEs within the chromophore. To achieve this, a number of differently labelled samples were prepared to test the applicability of the various assignment and distance measurement strategies. In measurements with and without light,1H and 13C chemical shifts of the retinal in the native protein were partially assigned for both the dark- and the light-adapted states. Additionally, samples with residue-specific1H amino acids and/or retinal in an otherwise deuterated protein were prepared to measure the distances between either two kinds of amino acids or between individual amino acids and the retinal moiety. With the observation of NOE within the bound retinal and between retinal and its neighbouring aminoacids, an important step towards the elucidation of distance constraints in the binding pocket of the proton pump is made

  8. Time-resolved laser studies on the proton pump mechanism of bacteriorhodopsin. Progress report, January 31,1991--February 1, 1992

    Energy Technology Data Exchange (ETDEWEB)

    El-Sayed, M.A.

    1991-12-31

    The research work carried out in 1991 can be classified as follows: (1) Work on the nature of the binding site of Eu{sup 3+} in which a fluorescence technique was used to determine the binding equilibrium constant from the concentration of the free Eu{sup 3+} in equilibrium with the bound ions. (2) The mechanism of the slow deprotonation process of bacteriorhodopsin during its photocycle from the observed temperature and pH dependence of its kinetics. (3) Using the circular dichroism spectrum of bR and its perturbed forms to examine the nature of the primary process as well as the origin of the non-exponential kinetic behavior of its photocycle. (4) Studies of bacteriorhodopsin mutants to identify the important amino acids that are part of the reaction coordinate of the deprotonation process as well as to assign the species that are important in giving rise to UV transient absorption whose origin was controversial.

  9. Protein changes associated with reprotonation of the Schiff base in the photocycle of Asp96-->Asn bacteriorhodopsin. The MN intermediate with unprotonated Schiff base but N-like protein structure

    Science.gov (United States)

    Sasaki, J.; Shichida, Y.; Lanyi, J. K.; Maeda, A.

    1992-01-01

    The difference Fourier transform infrared spectrum for the N intermediate in the photoreaction of the light-adapted form of bacteriorhodopsin can be recorded at pH 10 at 274 K (Pfefferle, J.-M., Maeda, A., Sasaki, J., and Yoshizawa, T. (1991) Biochemistry 30, 6548-6556). Under these conditions, Asp96-->Asn bacteriorhodopsin gives a photoproduct which shows changes in protein structure similar to those observed in N of wild-type bacteriorhodopsin. However, decreased intensity of the chromophore bands and the single absorbance maximum at about 400 nm indicate that the Schiff base is unprotonated, as in the M intermediate. This photoproduct was named MN. At pH 7, where the supply of proton is not as restricted as at pH 10, Asp96-->Asn bacteriorhodopsin yields N with a protonated Schiff base. The Asn96 residue, which cannot deprotonate as Asp96 in wild-type bacteriorhodopsin, is perturbed upon formation of both MN at pH 10 and N at pH 7. We suggest that the reprotonation of the Schiff base is preceded by a large change in the protein structure including perturbation of the residue at position 96.

  10. Recent Advances in the Field of Bionanotechnology: An Insight into Optoelectric Bacteriorhodopsin, Quantum Dots, and Noble Metal Nanoclusters

    Directory of Open Access Journals (Sweden)

    Christopher Knoblauch

    2014-10-01

    Full Text Available Molecular sensors and molecular electronics are a major component of a recent research area known as bionanotechnology, which merges biology with nanotechnology. This new class of biosensors and bioelectronics has been a subject of intense research over the past decade and has found application in a wide variety of fields. The unique characteristics of these biomolecular transduction systems has been utilized in applications ranging from solar cells and single-electron transistors (SETs to fluorescent sensors capable of sensitive and selective detection of a wide variety of targets, both organic and inorganic. This review will discuss three major systems in the area of molecular sensors and electronics and their application in unique technological innovations. Firstly, the synthesis of optoelectric bacteriorhodopsin (bR and its application in the field of molecular sensors and electronics will be discussed. Next, this article will discuss recent advances in the synthesis and application of semiconductor quantum dots (QDs. Finally, this article will conclude with a review of the new and exciting field of noble metal nanoclusters and their application in the creation of a new class of fluorescent sensors.

  11. Application of polymer films based on bacteriorhodopsin and its analogs for low-light-level imaging systems

    Science.gov (United States)

    Korchemskaya, Elena Y.; Soskin, Marat S.; Stepanchikov, Dmitriy A.; Djukova, T. V.; Druzhko, Anna B.; Vsevolodov, Nicolai N.

    1995-03-01

    In recent years polymer films based on bacteriorhodopsin (BR) have attracted a lot of attention in the area of optical imaging systems. The high photosensitivity of these films allows the processing of low-power optical signals (several mW/cm2 CW gas laser irradiation). Spatial resolution does not fall below 5000 lines/mm, photoresponse time is 50 microsecond(s) and images can be recorded and erased over million cycles. Polymer film with BR combine a dynamic recording with optical image processing. The characteristics of anisotropically-saturating nonlinearity of polymer films with BR allow a suppression of the background with greater intensity than usable signal intensity of be performed. Low saturation intensity of the polymer films with BR allows the operation of the polarization of low-intensity signals to be realized. Nonlinear photoresponse of the high photosensitivity BR genetic variant Asp96-Glu is studied in this work too. We hope that the polymer films based on BR and its analogs will find potential use precisely in the medical low- light-level imaging systems.

  12. All-optical switching in bacteriorhodopsin based on M state dynamics and its application to photonic logic gates

    Science.gov (United States)

    Singh, Chandra Pal; Roy, Sukhdev

    2003-03-01

    All-optical switching has been theoretically analyzed in bacteriorhodopsin (bR) based on nonlinear intensity induced excited state absorption of the M state. The transmission of a cw probe laser beam at 410 nm corresponding to the peak absorption of M state through a bR film is switched by a pulsed pump laser beam at 570 nm that corresponds to the maximum initial B state absorption. The switching characteristics have been numerically simulated using the rate equation approach considering all the six intermediate states (B, K, L, M, N and O) in the bR photocycle. The switching characteristics are shown to be sensitive to various parameters such as the pump pulse width, pump intensity, life time of the M state, thickness of the film and absorption cross-section of the B-state at probe wavelength ( σBp). It has been shown that the probe laser beam can be completely switched off (100% modulation) by the pump laser beam at relatively low pump powers, for σBp=0. The switching characteristics have also been used to theoretically design all-optical NOT, OR, AND and the universal NOR and NAND logic gates with two pulsed pump laser beams using the six state model.

  13. Structural Transition of Bacteriorhodopsin Is Preceded by Deprotonation of Schiff Base: Microsecond Time-Resolved X-Ray Diffraction Study of Purple Membrane

    OpenAIRE

    Oka, Toshihiko; Inoue, Katsuaki; Kataoka, Mikio; Yagi, Naoto

    2004-01-01

    The structural changes in the photoreaction cycle of bacteriorhodopsin, a light-driven proton pump, was investigated at a resolution of 7 Å by a time-resolved x-ray diffraction experiment utilizing synchrotron x rays from an undulator of SPring-8. The x-ray diffraction measurement system, used in coupling with a pulsed YAG laser, enabled us to record a diffraction pattern from purple membrane film at a time-resolution of 6 μs over the time domain of 5 μs to 500 ms. In the time domain, the fun...

  14. Photochromic Bacteriorhodopsin Mutant with High Holographic Efficiency and Enhanced Stability via a Putative Self-Repair Mechanism

    Science.gov (United States)

    2015-01-01

    The Q photoproduct of bacteriorhodopsin (BR) is the basis of several biophotonic technologies that employ BR as the photoactive element. Several blue BR (bBR) mutants, generated by using directed evolution, were investigated with respect to the photochemical formation of the Q state. We report here a new bBR mutant, D85E/D96Q, which is capable of efficiently converting the entire sample to and from the Q photoproduct. At pH 8.5, where Q formation is optimal, the Q photoproduct requires 65 kJ mol-1 of amber light irradiation (590 nm) for formation and 5 kJ mol-1 of blue light (450 nm) for reversion, respectively. The melting temperature of the resting state and Q photoproduct, measured via differential scanning calorimetry, is observed at 100 °C and 89 °C at pH 8.5 or 91 °C and 82 °C at pH 9.5, respectively. We hypothesize that the protein stability of D85E/D96Q compared to other blue mutants is associated with a rapid equilibrium between the blue form E85(H) and the purple form E85(−) of the protein, the latter providing enhanced structural stability. Additionally, the protein is shown to be stable and functional when suspended in an acrylamide matrix at alkaline pH. Real-time photoconversion to and from the Q state is also demonstrated with the immobilized protein. Finally, the holographic efficiency of an ideal thin film using the Q state of D85E/D96Q is calculated to be 16.7%, which is significantly better than that provided by native BR (6–8%) and presents the highest efficiency of any BR mutant to date. PMID:24498928

  15. Nonlinear Optical Properties of Bacteriorhodopsin and Retinal Chromophores and Their Applications for Optical Information Storage and Processing.

    Science.gov (United States)

    Chen, Zhongping

    Retinal, a conjugated polyene, plays a crucial role in biology. Both the visual pigments and the energy transducing protein, bacteriorhodopsin (BR) have a form of retinal as their chromophores. Because visual excitation and energy transduction in these systems is initiated by the promotion of retinal to an excited electronic state, information about the excited-state structure of retinal and the effect of chromophore/protein interactions on this structure are essential to understanding the functions of these systems. In this thesis, surface second harmonic (SH) generation is used to measure the light-induced dipole moment changes of a series of retinal derivatives that were designed and synthesized to model specific components of chromophore/protein interactions. In addition, we report an in situ probe of the dipole moment change of the retinal chromophore bound in BR by SH generation from oriented purple membranes. The dipole moment changes of various forms of BR, including light-adapted, dark-adapted, blue, and acid purple membrane, were measured and compared. These results, combined with the results from model compounds, elucidate the effects of the chromophore/protein interactions on light-induced charge redistribution and give insight on the fundamental nature of light excitation and energy storage in SR and rhodopsin. Furthermore, the dependence of the molecular hyperpolarizability of the conjugated molecules on donor/acceptor strength, protonation, conjugate length, planarity, and nonconjugate charges is investigated. Our study shows for the first time that nonconjugated charges have a very large effect on the nonlinear optical properties of conjugated molecules. BR has interesting photochromic characteristics, very large optical nonlinearities, and a unique optoelectrical property where the polarity of the photovoltage depends on both its photochromic state and the excitation wavelength. These unique characteristics coupled with its high stability make BR

  16. Probing bacteriorhodopsin photochemistry with nonlinear optics. Comparing the second harmonic generation of bR and the photochemically induced intermediate K

    Energy Technology Data Exchange (ETDEWEB)

    Bouevitch, O.; Lewis, A. [Hebrew Univ., Jerusalem (Israel); Sheves, M. [Weizmann Inst. of Science, Rehevot (Israel)

    1995-06-29

    The nonlinear optical properties of the bacteriorhodopsin chromophore in the bR568 and K states are investigated by second harmonic generation. The comparison of amplitudes and phases of the second-order nonlinear optical polarizabilities of the retinal chromophore in the two states has revealed a noticeable increase of the induced dipole of the retinal as a result of the bR568 $YLD K transition. The results have been explained in terms of recent theoretical understandings of the nonlinear optical properties of polyenes. Within the context of these understandings we have discussed the molecular origins of the light-induced color changes and the possible mechanism of photon energy storage observed in this protein. 54 refs., 8 figs., 3 tabs.

  17. A pKa calculation of residues in a proton pump, bacteriorhodopsin, from structures determined by electron crystallography.

    Science.gov (United States)

    Mitsuoka, Kaoru

    2014-11-01

    Bacteriorhodopsin (bR) is a light-driven proton pump, which is a membrane protein found in halophilic archeae like Halobacterium salinarum and in eubacteria [1]. When the covalently bound retinal chromophore absorbs the light energy, it changes the conformation from all-trans to 13-cis. This configuration change initiates ion translocation across the cell membrane and a proton moves from inside to outside of the cell. The bR molecules are forming two-dimensional crystals on the membranes of halophilic archeae, and therefore the atomic model of bR was first determined by electron crystallography. The determined structure can be used to determine the pKa values, through which the charge states of ionizable residues in bR determine their pH-dependent properties. The pH-dependent properties are crucial for proton translocation from ionizable residues or to ionizable residues. Detection of the intermediate states of the reaction cycle (photocycle) produced spectroscopic information, which can predict the ionization state of the ionozable residues. In the transition from the L intermediate to the M intermediate, it is known that a proton moves from the Shiff base on the retinal chromophore to Asp85, while a proton is released to the extracellar side from proton-releasing groups including Glu194 and Glu204. Experimentally the pKa value of the proton release is determined to be about 9.7, while the pKa value of Asp85 was measured to change from 2.6 to 7.5 by the proton release from the proton-releasing groups [2]. Here we used the PROPKA program [3] to calculate the pKa values of Asp85 and the proton-releasing groups from the structures at pH 5.5 and at pH 10.0 determined by electron crystallography. The calculation showed that the pKa value of Asp85 changes from 5.3 to 6.1, which qualitatively show the similar changes with the measured difference. The largest change between the structures is the shift of Arg82 by the proton release from the proton-releasing groups

  18. FTIR Studies of Internal Water Molecules of Bacteriorhodopsin: Structural Analysis of Halide-bound D85S and D212N Mutants in the Schiff Base Region

    Science.gov (United States)

    Shibata, Mikihiro; Kandori, Hideki

    2007-12-01

    Bacteriorhodopsin (BR), a membrane protein found in Halobacterium salinarum, functions as a light-driven proton pump. The Schiff base region has a quadropolar structure with positive charges located at the protonated Schiff base and Arg82, and counterbalancing negative charges located at Asp85 and Asp212 (Figure 1A). It is known that BR lacks a proton-pumping activity if Asp85 or Asp212 is neutralized by mutation. On the other hand, binding of C1- brings different effects for pumping functions in mutants at D85 and D212 position. While C1--bound D85T and D85S pump C1-, photovoltage measurements suggested that C1--bound D212N pumps protons at low pH. In this study, we measured low-temperature FTIR spectra of D85S and D212N containing various halides to compare the halide binding site of both proteins. In the case of D85S, the N-D stretching vibrations of the Schiff base were halide-dependent. This result suggests that the halide is a hydrogen-bond acceptor of the Schiff base, being consistent with the X-ray crystal structure. On the other hand, no halide dependence was observed for vibrational bands of the retinal skeleton and the Schiff base in the D212N mutant. This result suggests that the halide does not form a hydrogen bond with the Schiff base directly, unlike the mutation at D85 position. Halide-dependent water bands in the Schiff base region also differ between D85S and D212N. From these results, halide binding site of both proteins and role of two negative charges in BR will be discussed.

  19. Second-harmonic generation of biological interfaces: probing the membrane protein bacteriorhodopsin and imaging membrane potential around GFP molecules at specific sites in neuronal cells of C. elegans

    Science.gov (United States)

    Lewis, Aaron; Khatchatouriants, Artium; Treinin, Millet; Chen, Zhongping; Peleg, Gadi; Friedman, Noga; Bouevitch, Oleg; Rothman, Zvi; Loew, Leslie; Sheres, Mordechai

    1999-07-01

    Second-harmonic generation (SHG) is applied to problems of probing membrane proteins and functionally imaging around selective sites and at single molecules in biological membranes. The membrane protein bacteriorhodopsin (bR) has been shown to have large second-harmonic (SH) intensities that are modulated by protein/retinylidene chromophore interactions. The nonlinear optical properties of model compounds, which simulate these protein chromophore interactions in retinal proteins, are studied in this work by surface SHG and by hyper-Rayleigh scattering. Our results indicate that non-conjugated charges and hydrogen bonding effects have a large effect on the molecular hyperpolarizability of the retinal chromophore. However, mbR, the model system studies suggest that polarizable amino acids strongly affect the vertically excited state of the retinylidene chromophore and appear to play the major role in the observed protein enhancement (>50%) of the retinylidene chromophore molecular hyperpolarizability and associated induced dipole. Furthermore, the data provide insights on emulating these interactions for the design of organic nonlinear optical materials. Our studies have also led to the development of dyes with large SH intensities that can be embedded in cell membranes and can functionally image membrane potential. Single molecules of such dyes in selected single molecular regions of a cell membrane have been detected. SHG from green fluorescent protein (GFP) selectively expressed in concert with a specific protein in neuronal cells in a transgenic form of the worm C. elegans is also reported. The membrane potential around the GFP molecules expressed in these cells has been imaged with SHG in live animals.

  20. Isolation of a new Pseudomonas halophila strain possess bacteriorhodopsin-like protein by a novel method for screening of photoactive protein producing bacteria.

    Science.gov (United States)

    Fanaei, Maryam; Emtiazi, Giti

    2014-02-01

    Bacteriorhodopsin (bR) is a transmembrane protein deposited in the purple membrane of Halobacterium salinarum which absorbs energy from photons to create a photo-induced proton gradient across the membrane. A bR molecule can be considered as a natural solar device transforming light into other types of energy and therefore is of interest for a wide range of applications including two and three-dimensional memory storage, optical data processing, artificial cells, holographic media, the artificial retina and photo sensor devices. H. salinarum is a slow-growing, halophilic Archaea present in red salt waters. The present study introduces a novel bR-like pigment from a new strain of Pseudomonas halophila (with registered accession number KC959570 in the NCBI databank) which has a very significant degree of light-dependent activity. This is the first report on the presence of functional bR-like protein in the Pseudomonas family. The isolate is a fast-growing, halophilic bacterium and is comparable with other photoactive protein producer microorganisms. Also, in the present study a novel isolation method for screen light-stimulating protein producing microorganisms is introduced. For this purpose 2,3,5-triphenyltetrazolium chloride (TTC) was employed for the first time as an artificial hydrogen acceptor in the proton-transfer processes. The TTC test is an easy and susceptible method for estimating hydrogen production during the proton transport process. This is the first report of the use of TTC for photo activity measurement and selection of bacteria containing light dependent proteins. PMID:24002576

  1. Nonlinear Optical Characteristic Measurement of Chemically Enhanced Bacteriorhodopsin Film Using Z-scan Technology%利用Z扫描技术测量菌紫质膜的非线性光学参数

    Institute of Scientific and Technical Information of China (English)

    徐军; 何俊发; 侯素霞; 胡坤生; 张亦南

    2000-01-01

    本文讨论一种有发展前景的光敏生物材料-细菌视紫红质(菌紫质),采用化学增强法制备菌紫质膜,利用Z扫描技术测量了菌紫质膜的非线性光学参数,结果表明菌紫质具有大的非线性光学系数,在光信息处理领域有十分广泛的应用.%We present a unique biological material that exhibits interesting photochromic characteristics and important optoelectric properties. This distinctive material is related to the visual pigment rhodopsin and is called bacteriorhodopsin (bR). In this paper the films of the chemically enhanced bacteriorhodopsin are prepared and the effective nonlinearity n2 of the film is measured with the Z-scan Technology. Anomalous absorption at three wavelengths is observed. Large optical nonlinearities of bR should be useful for both information storage and compulation.

  2. Quadratic electro-optic effects in bacteriorhodopsin: Measurement of γ(-ω;0,0,ω) in dried gelatin thin films

    Science.gov (United States)

    Yamazaki, Mikio; Goodisman, Jerry; Birge, Robert R.

    1998-04-01

    Quadratic electro-optic effects (dc or low frequency Kerr effect) of bacteriorhodopsin dispersed in dried gelatin thin films are examined in the near resonance region at three wavelengths: 633, 647, and 676 nm. The films show relatively large quadratic electro-optic effects compared to other molecular dispersed systems. The purple membrane is fixed within the polymerized gelatin matrix, and we show that the electronic contribution to γ dominates over possible orientational contributions. At 676 nm, the quadratic electro-optic coefficient s1133(-ω;0,0,ω) is 6.7×10-20m2/V2 and the third order nonlinear susceptibility χ1133(3)(-ω;0,0,ω) is 7.0×10-13cm4 statCoulomb-2, with both values obtained for a protein concentration of 6.9×1018cm-3. The orientationally averaged second molecular hyperpolarizability determined from the quadratic electro-optic coefficients at 676 nm assuming an Onsager ellipsoidal local field factor is (10.8±5.1)×10-32 cm7 statCoulomb-2 [(1.34±0.63)×10-56 F3 m4 C-2]. The value increases roughly tenfold when the probe wavelength is decreased to 633 nm. The behavior of γ(-ω;0,0,ω), when fit to a two-state model, predicts that γ(-ω;0,0,ω) is strongly enhanced via type III processes. Thus, the magnitude of γ(-ω;0,0,ω) is dominated by a term (Δμ102×μ102)/(ω10-ω)3, where Δμ10 is the change in dipole moment, μ10 is the transition moment, and ω10 is the transition energy of the lowest-lying allowed 1Bu*+-like π,π* state. We calculate that Δμ10 is 12.8±1.2 D, in good agreement with previous Stark and two-photon experimental values. Time-dependent Hartree-Fock methods based on the MNDO Hamiltonian yield reasonable agreement with experiment, underestimating γ(-ω;0,0,ω) by factors of only 2-4, with the error increasing as the frequency approaches resonance.

  3. Incorporation of the dopamine D2L receptor and bacteriorhodopsin within bicontinuous cubic lipid phases. 2. Relevance to in meso crystallization of integral membrane proteins in novel lipid systems

    Energy Technology Data Exchange (ETDEWEB)

    Conn, Charlotte E.; Darmanin, Connie; Sagnella, Sharon M.; Mulet, Xavier; Greaves, Tamar L.; Varghese, Joseph N.; Drummond, Calum J.

    2014-09-24

    The dopamine D2 long (D2L) receptor and bacteriorhodopsin (bR), which are integral membraneproteins, have been incorporated within bicontinuous cubic mesophases formed by the lipids anandamide and H-farnesoyl monoethanolamide, which have been specifically investigated by us for use as in mesocrystallization media. We show that the incorporated membraneprotein affects the structure of the cubic phases with the particular effect observed dependent on the geometry of the underlying cubic phase. The results are complementary to those obtained in Part 1 of this series, where we demonstrated that the structural effects observed depend on the structure of the membraneprotein. Importantly protein concentrations commonly used for crystallization can destroy the cubic phase matrix, particularly where there is a large discrepancy between the hydrophilic and the hydrophobic spans of the membraneprotein, and the hydrophilic and hydrophobic domain sizes of the cubic phase.

  4. Determination of the Waist Position of a Gaussian Beam by Bacteriorhodopsin Film%由细菌视紫红质测定高斯光束的束腰位置

    Institute of Scientific and Technical Information of China (English)

    陈桂英; 郭宗霞; 张春平; 田建国; Q.W.Song; Mingchien Huang

    2004-01-01

    Relation between transmitted intensity of bacteriorhodopsin(bR) film and the incident intensity was tested.A new method of determining waist position of a Gaussian beam passing an optical system was proposed by the nonlinear transmission of the bR film.The measured results are in agreement with the calculated results based on the parameters of the Guassian beam.%介绍并测量了细菌视紫红质(bR)的透过光强随入射光强的变化特性,并提出利用bR的非线性透过特性测定高斯光束的束腰位置,测量结果与利用已知的高斯光束参数所计算的结果相一致.

  5. 细菌视紫红质激子的饱和密度及激子长度的研究%Study on Saturation Density and Length of Excitons in Bacteriorhodopsin

    Institute of Scientific and Technical Information of China (English)

    黄燕萍; 陈树德; 王祖赓; 李庆国

    2001-01-01

    The three-energy model simulated with photocycle characteristicsis used to explain bacteriorhodopsin’s 3-order nonlinear susceptibility and its time response, which was measured with four-wave mixing technique. The saturation density and length of excitons in bacteriorhodopsin are obtained with fitting to the phase-space filling model. The possible mechanism for generating the 3rd nonlinear optical susceptibility χ(3) and response time are discussed.%对光能转换生物分子细菌视紫红质(bR)作前向立体简并四波混频实验所测得的三阶非线性电极化率和它们的时间响应,用光循环结构模拟的三能级模型予以解释,并用位相空间充满理论模拟得到激子饱和密度和激子长度。

  6. Nonlinear phase contrast using a bacteriorhodopsin film

    Science.gov (United States)

    Iturbe Castillo, Marcelo D.; Sanchez-de-la-Llave, J. D.; Ramos Garcia, Ruben; Tepichin-Rodriguez, Eduardo; Olivos-Perez, L. I.

    2002-11-01

    In this paper we demonstrate a novel phase contrast system that employs a BR film. Since the filter is optically induced by the Fourier transform of the phase object, no alignment is necessary at the filter plane making it extremely robust. Due to the optical properties of BR films the phase filter can be induced with low light intensity levels. The material response allows operation at video frame rates, processing of high spatial resolution objects, and the use of relatively inexpensive laser sources. Such characteristics and the fact that BR films can be produced at a low cost makes the system simple to implement, relatively inexpensive and extremely robust. The effects of varying the illuminating area beyond the phase object area and filter saturation are also analyzed.

  7. Holographic particle image velocimetry using Bacteriorhodopsin

    NARCIS (Netherlands)

    Koek, W.D.

    2006-01-01

    To gain better insight into the behaviour of turbulent flow there is a demand for a practical measurement instrument to perform three-dimensional flow measurements. Holography is a three-dimensional imaging technique, and as such is ideally suited for this purpose. Because flow media (such as water

  8. Spectral Signatures of the Pentagonal Water Cluster in Bacteriorhodopsin

    International Nuclear Information System (INIS)

    The exchange of protons between basic and acidic groups within proteins often involves transient protonation of amino acids and water molecules embedded in the protein matrix. One of the best studied proteins in this respect is Bacteriorohodopsin (BR), which works in the membrane of Halobacterium salinarium as a light-driven proton pump. The pumping process is triggered in the initial bR state by a photon absorption of an all-trans retinylidene chromophore, which is linked via a protonated Schiff base (pRSB) to the sidechain of Lys216. The subsequent photocycle comprises a series of intermediate states J, K, L, M, N and O, which are characterized by conformational and absorbance changes of the chromophore accompanying several elementary proton transfer processes. Upon completion of the photocycle one net proton has been transferred from the cyctoplasmic to the extracellular side against the proton gradient across the membrane. These proton exchange reactions can be monitored by time resolved infrared (IR) spectroscopy of the BR wild type and site specific mutants, which allow the localization of absorbance changes within the protein. Furthermore, these measurements have revealed the fundamental importance of internal water molecules in these processes as supported by recent large-scale QM/MM molecular dynamics studies of anharmonic IR spectra

  9. Integrated optical devices using bacteriorhodopsin as active nonlinear optical material

    OpenAIRE

    Dér, A; Fábián, L.; Valkai, S.; Wolff, E.; Ramsden, Jeremy J.; Ormos, P.

    2006-01-01

    Coupling of optical data-processing devices with microelectronics, telecocommunication and sensory functions, is among the biggest challenges in molecular electronics. Intensive research is going on to find suitable nonlinear optical materials that could meet the demanding requirements of optoelectronic applications, especially regarding high sensitivity and stability. In addition to inorganic and organic crystals, biological molecules have also been considered for use in in...

  10. First hyperpolarizability of bacteriorhodopsin, retinal and related molecules revisited

    Science.gov (United States)

    Ghosh, Sampa; Ranjini, Arumugam Sri; Pandey, Ravindra; Das, Puspendu Kumar

    2009-06-01

    The previously reported β values of BR and retinal based chromophores were very high but subsequent measurements found them to be much less. We have found that the β values of these compounds do not vary so much with experimental conditions as with the method of analysis. Hyper-Rayleigh scattering measurements at 1543 and 1907 nm produce more realistic β values close to the intrinsic (static) hyperpolarizability, β0 which for BR is still very high (275 × 10 -30 esu). The optical nonlinearity of BR arises entirely due to the protonated retinal Schiff Base (PRSB) which in its isolated form has the same intrinsic hyperpolarizability as that of the protein.

  11. Real-time holography on bacteriorhodopsin-based materials

    Science.gov (United States)

    Taranenko, Victor B.

    1998-09-01

    The main properties and mechanisms of photoresponse of the bacteriohodopsin-based materials are presented. Fields of their potential applications in the real-time holography and nonlinear optics are discussed.

  12. Reusable holographic velocimetry system based on polarization multiplexing in Bacteriorhodopsin

    NARCIS (Netherlands)

    Koek, W.D.; Chan, V.S.S.; Ooms, T.A.; Bhattacharya, N.; Westerweel, J.; Braat, J.J.M.

    2005-01-01

    We present a novel holographic particle image velocimetry (HPIV) system using a reversible holographic material as the recording medium. In HPIV the three-dimensional flow field throughout a volume is detected by adding small tracer particles to a normally transparent medium. By recording the partic

  13. Spectral Signatures of the Pentagonal Water Cluster in Bacteriorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Baer, M; Mathias, G; Kuo, I W; Tobias, D J; Mundy, C J; Marx, D

    2008-07-25

    The exchange of protons between basic and acidic groups within proteins often involves transient protonation of amino acids and water molecules embedded in the protein matrix. One of the best studied proteins in this respect is Bacteriorohodopsin (BR), which works in the membrane of Halobacterium salinarium as a light-driven proton pump. The pumping process is triggered in the initial bR state by a photon absorption of an all-trans retinylidene chromophore, which is linked via a protonated Schiff base (pRSB) to the sidechain of Lys216. The subsequent photocycle comprises a series of intermediate states J, K, L, M, N and O, which are characterized by conformational and absorbance changes of the chromophore accompanying several elementary proton transfer processes. Upon completion of the photocycle one net proton has been transferred from the cyctoplasmic to the extracellular side against the proton gradient across the membrane. These proton exchange reactions can be monitored by time resolved infrared (IR) spectroscopy of the BR wild type and site specific mutants, which allow the localization of absorbance changes within the protein. Furthermore, these measurements have revealed the fundamental importance of internal water molecules in these processes as supported by recent large-scale QM/MM molecular dynamics studies of anharmonic IR spectra.

  14. Mass spectrometric analysis of integral membrane proteins: application to complete mapping of bacteriorhodopsins and rhodopsin.

    OpenAIRE

    Ball, L. E.; Oatis, J. E.; Dharmasiri, K.; Busman, M.; Wang, J.; Cowden, L. B.; Galijatovic, A.; N. Chen; Crouch, R K; Knapp, D R

    1998-01-01

    Integral membrane proteins have not been readily amenable to the general methods developed for mass spectrometric (or internal Edman degradation) analysis of soluble proteins. We present here a sample preparation method and high performance liquid chromatography (HPLC) separation system which permits online HPLC-electrospray ionization mass spectrometry (ESI-MS) and -tandem mass spectrometry (MS/MS) analysis of cyanogen bromide cleavage fragments of integral membrane proteins. This method has...

  15. Optical nonlinearity of pure bacteriorhodopsin Langmuir-Blodgett films derived from multi-wave mixing

    Science.gov (United States)

    Liu, S. H.; Du, Weichong

    1993-10-01

    We report an observation of optical phase conjugate and high-order diffractions from degenerate multi-wave mixing in LB films of pure purple membrane for the first time. The saturated absorption intensity and the saturated nonlinear refractive index of the LB films have been estimated to be 0.42 W/cm2, and 5×10-2 cm2/W, respectively. The typical response time of its nonlinearity is about several milliseconds.

  16. Orientation of a bacteriorhodopsin thin film deposited by dip coating technique and its chiral SHG as studied by SHG interference technique

    Science.gov (United States)

    Yamada, Toshiki; Haruyama, Yoshihiro; Kasai, Katsuyuki; Terui, Toshifumi; Tanaka, Shukichi; Kaji, Takahiro; Kikuchi, Hiroshi; Otomo, Akira

    2012-03-01

    We show that by observing SHG interference bR thin films prepared by a simple dip coating technique have a polar orientation with C∞ symmetry. The SHG interference measurements were performed under various input and output polarization combinations at different incident angles or under the rotation of the quarter-wave retardation plate at specific incident angles. The interference patterns provide us with insight into the characteristics of non-vanishing nonlinear optical coefficients including chiral components. Abundant information can be obtained by observing SHG interference by using two chiral SH active films.

  17. PHOTONIC SWITCHES BASED ON BACTERIORHODOPSIN%细菌视紫红质光开关特性实验研究

    Institute of Scientific and Technical Information of China (English)

    田燕宁; 冯晓强; 侯洵; 陈烽

    2001-01-01

    介绍实现光子开关材料BR分子的重要性。理论上分析BR分子材料光学特性,实验观测BR分子材料在400nm和632nm光照射下的相互抑制作用,最后分析BR的光子开关特性。%The importance of BR which is used in achieving photonic switches is summarized simply.The principle and ways how to realized photonic switches based on BR are analyzed in detail.Under illuminatiing by two beams at 568nm and 412nm,the transmission of two beams is moduleted by BR in a complementary fashion.Then,this paper shows the experiments of photonic switches.The datas from experiments are analyzed finally.

  18. Third Order Nonlinear Optical Properties of Bacteriorhodopsin Langmuir-Blodgett Multilayer Film%菌紫质LB膜的三阶非线性光学性能的研究

    Institute of Scientific and Technical Information of China (English)

    祁春媛; 刘康俊; 黄燕萍; 钱士雄

    2006-01-01

    在ITO导电玻璃上,制作38层Z型细菌视紫红质(bR)的LB膜;控制平均转移比在0.93以上.测量了这个LB膜的紫外-可见吸收谱.利用Z扫描技术在输出飞秒激光,波长为400nm和800nm处对菌紫质LB膜的三阶非线性光学性能进行了研究.在800nm处,它的三阶非线性光学极化率为10-9esu, 而在400nm 处为10-8esu.这表明菌紫质LB膜在非线性光学器件方面具有潜在的应用前景.

  19. 细菌视紫红质能化态时表面电位的非线性光学机制%Study on Nonlinear Optical Mechanism of Surface Potential in Energized Bacteriorhodopsin

    Institute of Scientific and Technical Information of China (English)

    黄燕萍; 沈珊雄; 李琳; 李珊珊; 黄立; 李庆国

    2003-01-01

    用荧光标记物1,8-ANS与细菌视紫红质结合,测得紫膜细菌视紫红质在能化态时的表面电位远大于非能化态时的对应值.在细菌视紫红质分子的激光四波混频实验中,应用激子表象理论,获得了紫膜能化态时的激子饱和密度和激子长度,说明在bR-ANS络合物中,细菌视紫红质中色氨酸残基对ANS的激发能量转移效率提高,能化态时表面电荷密度增加,从而使非辐射共振转移变为激子转移,也证明了紫膜能化态时表面电位的非线性光学机制.

  20. Real-time UV-visible spectroscopy analysis of purple membrane-polyacrylamide film formation taking into account Fano line shapes and scattering.

    Directory of Open Access Journals (Sweden)

    María Gomariz

    Full Text Available We theoretically and experimentally analyze the formation of thick Purple Membrane (PM polyacrylamide (PA films by means of optical spectroscopy by considering the absorption of bacteriorhodopsin and scattering. We have applied semiclassical quantum mechanical techniques for the calculation of absorption spectra by taking into account the Fano effects on the ground state of bacteriorhodopsin. A model of the formation of PM-polyacrylamide films has been proposed based on the growth of polymeric chains around purple membrane. Experimentally, the temporal evolution of the polymerization process of acrylamide has been studied as function of the pH solution, obtaining a good correspondence to the proposed model. Thus, due to the formation of intermediate bacteriorhodopsin-doped nanogel, by controlling the polymerization process, an alternative methodology for the synthesis of bacteriorhodopsin-doped nanogels can be provided.

  1. Dynamic holography in optical pattern recognition

    Science.gov (United States)

    Rajbenbach, Henri J.

    1994-03-01

    This paper reviews the basic principles, physical processes, and most recent demonstrations of optical correlators using dynamic holographic techniques in nonlinear media such as bulk photorefractives (PR), thin multiple quantum wells (MQW), and bacteriorhodopsin (BR) films.

  2. Light energy transduction by the purple membrane of halophilic bacteria; Proceedings of the Symposium, San Francisco, Calif., June 6, 1976

    Science.gov (United States)

    1977-01-01

    Several aspects of bacteriorhodopsin, the retinal protein component of the purple membranes of Halobacterium halobium, are discussed. Structural studies are presented. Photochemical properties of the protein complex and of its chromophore are described. Proton translocation of bacteriorhodopsin is compared to that of a protein from a thermophilic bacterium. Ionophore activity of bacteriorhodopsin is considered with attention to conformational changes, light dependency, and electrical potential. Amino acid transport is also examined and the light-energy budget is investigated. Bacteriorhodopsin is of interest because of its similarity to rhodopsin, which plays a major role in mammalian vision, and also because its attainability and distinctive characteristics will facilitate studies of certain bacterial physiological functions, such as ion transport and membrane organization.

  3. Design and modeling of a light powered biomimicry micropump

    Science.gov (United States)

    Sze, Tsun-kay Jackie; Liu, Jin; Dutta, Prashanta

    2015-06-01

    The design of compact micropumps to provide steady flow has been an on-going challenge in the field of microfluidics. In this work, a novel micropump concept is introduced utilizing bacteriorhodopsin and sugar transporter proteins. The micropump utilizes light energy to activate the transporter proteins, which create an osmotic pressure gradient and drive the fluid flow. The capability of the bio inspired micropump is demonstrated using a quasi 1D numerical model, where the contributions of bacteriorhodopsin and sugar transporter proteins are taken care of by appropriate flux boundary conditions in the flow channel. Proton flux created by the bacteriorhodopsin proteins is compared with experimental results to obtain the appropriate working conditions of the proteins. To identify the pumping capability, we also investigate the influences of several key parameters, such as the membrane fraction of transporter proteins, membrane proton permeability and the presence of light. Our results show that there is a wide bacteriorhodopsin membrane fraction range (from 0.2 to 10%) at which fluid flow stays nearly at its maximum value. Numerical results also indicate that lipid membranes with low proton permeability can effectively control the light source as a method to turn on/off fluid flow. This capability allows the micropump to be activated and shut off remotely without bulky support equipment. In comparison with existing micropumps, this pump generates higher pressures than mechanical pumps. It can produce peak fluid flow and shutoff head comparable to other non-mechanical pumps.

  4. Photosensitive phosphoproteins in Halobacteria: regulatory coupling of transmembrane proton flux and protein dephosphorylation

    OpenAIRE

    1981-01-01

    A photoregulated reversible protein phosphorylation system controlled by the halobacterial rhodopsins was recently reported. The results presented in this paper identify the initial steps in the pathway from the absorption of light to the photoregulated protein phosphorylation and dephosphorylation reactions. Action spectrum, biochemical, and genetic analyses show that the proton pump bacteriorhodopsin mediates light-induced dephosphorylation of three photoregulated phosphoproteins. Light abs...

  5. Synthesis of ring-13C-labelled and ring-demethylated retinals

    International Nuclear Information System (INIS)

    Efficient synthetic schemes are described for the preparation of the required mono- and di-13C labelled retinals based on simple 13C labelled starting materials. Results from solid-state 13C-NMR spectroscopic studies of the various ring-13C labelled bacteriorhodopsins and rhodopsins are discussed. 404 refs.; 74 figs.; 16 tabs

  6. Protein-based integrated optical switching and modulation

    Science.gov (United States)

    Ormos, Pál; Fábián, László; Oroszi, László; Wolff, Elmar K.; Ramsden, Jeremy J.; Dér, András

    2002-05-01

    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.

  7. Expression and functioning of retinal-based proton pumps in a saltern crystallizer brine.

    Science.gov (United States)

    Oren, Aharon; Abu-Ghosh, Said; Argov, Tal; Kara-Ivanov, Eliahu; Shitrit, Dror; Volpert, Adi; Horwitz, Rael

    2016-01-01

    We examined the presence of bacteriorhodopsin and other retinal protein pigments in the microbial community of the saltern crystallizer ponds in Eilat, Israel, and assessed the effect of the retinal-based proton pumps on the metabolic activity. The biota of the hypersaline (~309 g salts l(-1)) brine consisted of ~2200 β-carotene-rich Dunaliella cells and ~3.5 × 10(7) prokaryotes ml(-1), most of which were flat, square or rectangular Haloquadratum-like archaea. No indications were obtained for massive presence of Salinibacter. We estimated a concentration of bacteriorhodopsin and bacteriorhodopsin-like pigments of 3.6 nmol l(-1). When illuminated, the community respiration activity of the brine samples in which oxygenic photosynthesis was inhibited by 3-(3-4-dichlorophenyl)-1,1-dimethylurea, decreased by 40-43 %. This effect was interpreted to be the result of competition between two energy yielding systems: the bacteriorhodopsin proton pump and the respiratory chain. The results presented have important implications for the interpretation of many published data on photosynthetic and respiratory activities in hypersaline environments. PMID:26507954

  8. Use of spin labels to study membrane proteins by high-frequency electron nuclear double resonance spectroscopy

    NARCIS (Netherlands)

    Orlinkskii, S.B.; Borovykh, I.V.; Zielke, V.; Steinhoff, H.J.

    2007-01-01

    The applicability of spin labels to study membrane proteins by high-frequency electron nuclear double resonance spectroscopy is demonstrated. With the use of bacteriorhodopsin embedded in a lipid membrane as an example, the spectra of protons of neighboring amino acids are recorded, electric field g

  9. Evolution of rhodopsin ion pumps in haloarchaea

    Directory of Open Access Journals (Sweden)

    Ford Doolittle W

    2007-05-01

    Full Text Available Abstract Background The type 1 (microbial rhodopsins are a diverse group of photochemically reactive proteins that display a broad yet patchy distribution among the three domains of life. Recent work indicates that this pattern is likely the result of lateral gene transfer (LGT of rhodopsin genes between major lineages, and even across domain boundaries. Within the lineage in which the microbial rhodopsins were initially discovered, the haloarchaea, a similar patchy distribution is observed. In this initial study, we assess the roles of LGT and gene loss in the evolution of haloarchaeal rhodopsin ion pump genes, using phylogenetics and comparative genomics approaches. Results Mapping presence/absence of rhodopsins onto the phylogeny of the RNA polymerase B' subunit (RpoB' of the haloarchaea supports previous notions that rhodopsins are patchily distributed. The phylogeny for the bacteriorhodopsin (BR protein revealed two discrepancies in comparison to the RpoB' marker, while the halorhodopsin (HR tree showed incongruence to both markers. Comparative analyses of bacteriorhodopsin-linked regions of five haloarchaeal genomes supported relationships observed in the BR tree, and also identified two open reading frames (ORFs that were more frequently linked to the bacteriorhodopsin gene than those genes previously shown to be important to the function and expression of BR. Conclusion The evidence presented here reveals a complex evolutionary history for the haloarchaeal rhodopsins, with both LGT and gene loss contributing to the patchy distribution of rhodopsins within this group. Similarities between the BR and RpoB' phylogenies provide supportive evidence for the presence of bacteriorhodopsin in the last common ancestor of haloarchaea. Furthermore, two loci that we have designated bacterio-opsin associated chaperone (bac and bacterio-opsin associated protein (bap are inferred to have important roles in BR biogenesis based on frequent linkage and co

  10. Processing of medical images using real-time optical Fourier processing

    International Nuclear Information System (INIS)

    Optical image processing techniques are inherently fast in view of parallel processing. A self-adaptive optical Fourier processing system using photoinduced dichroism in a bacteriorhodopsin film was experimentally demonstrated for medical image processing. Application of this powerful analog all-optical interactive technique for cancer diagnostics is illustrated with two mammograms and a Pap smear. Microcalcification clusters buried in surrounding tissue showed up clearly in the processed image. By playing with one knob, which rotates the analyzer in the optical system, either the microcalcification clusters or the surrounding dense tissue can be selectively displayed. Bacteriorhodopsin films are stable up to 140 deg. C and environmentally friendly. As no interference is involved in the experiments, vibration isolation and even a coherent light source are not required. It may be possible to develop a low-cost rugged battery operated portable signal-enhancing magnifier

  11. Imaging bacteriorhodopsinlike molecules of claretmembranes from Tibet halobacteria xz515 by atomic force microscope

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Halobacteria H. sp.xz 515 was isolated from a salt lake in Tibet. Although proton release-and-uptake across claret membrane is in reverse order compared to bacteriorhodopsin in purple membrane from Halobacterium Salinarum, and its efficiency of proton pump is much lower,AFM image shows that the molecules are still arranged in a two-dimensional hexagonal lattice of trimers. Primary structure of C- to G-helix of the archaerhodopsin shows that it has only 56% homology with bacteriorhodopsin. But the interactive amino acid residues at the interface between Band D-helixes are conserved. These amino acid residues are believed to play a significant role in the stability of protein oligomers.

  12. Dynamic nuclear polarization enhanced NMR at 187 GHz/284 MHz using an extended interaction Klystron amplifier

    OpenAIRE

    Watts, A; Kemp, T; Dannatt, HRW; Barrow, NS; Brown, SP; Newton, ME; Dupree, R.

    2016-01-01

    A Dynamic Nuclear Polarisation (DNP) enhanced solid-state Magic Angle Spinning (MAS) NMR spectrometer which uses a 187 GHz (corresponding to 1H NMR frequency of 284 MHz) Extended Interaction Klystron (EIK) amplifier as the microwave source is briefly described. Its performance is demonstrated for a biomolecule (bacteriorhodopsin), a pharmaceutical, and surface functionalised silica. The EIK is very compact and easily incorporated into an existing spectrometer. The bandwidth of the amplifier i...

  13. Investigation of structural change of purple membrane in storage by transmission electron microscope and atomic force microscope

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The structural change of purple membrane during storage has been investigated by means of transmission electron microscope and atomic force microscope. It is found that many liposomes have spontaneously evolved from the purple membrane sheets isolated three years ago. The membrane proteins on the liposomes, bacteriorhodopsin, are still presented as trimers in 2-D hexagonal structure, which is the same as that in natural cell membrane. However, the cytoplasmic surface of purple membrane faced outside on the liposomes.

  14. Cooperative long range protein-protein dynamics in Purple Membrane

    OpenAIRE

    Rheinstadter, Maikel; Schmalzl, Karin; Wood, Kathleen; Strauch, Dieter

    2008-01-01

    We present experimental evidence for a long-range protein-protein interaction in purple membrane (PM). The interprotein dynamics were quantified by measuring the spectrum of the acoustic phonons in the 2D bacteriorhodopsin (BR) protein lattice using inelastic neutron scattering. Phonon energies of about 1 meV were determined. The data are compared to an analytical model, and the effective spring constant for the interaction between neighboring protein trimers are determined to be k=53 N/m. Ad...

  15. Proteins as nano-machines: dynamics-function relations studied by neutron scattering

    International Nuclear Information System (INIS)

    A protein is a nano-machine whose molecular structure was selected by evolution to perform specific biological functions. Neutron spectroscopy is uniquely suited to provide experimental data on atomic motions in a protein under the influence of forces that maintain its stable and active molecular structure. Experiments are reviewed relating such a dynamics to soluble protein folding and stability in different environments, and to the activity of bacteriorhodopsin, a membrane protein nano-machine with light-driven proton pump activity

  16. Ionic Polymer Microactuator Activated by Photoresponsive Organic Proton Pumps

    OpenAIRE

    Khaled M. Al-Aribe; George K. Knopf; Amarjeet S. Bassi

    2015-01-01

    An ionic polymer microactuator driven by an organic photoelectric proton pump transducer is described in this paper. The light responsive transducer is fabricated by using molecular self-assembly to immobilize oriented bacteriorhodopsin purple membrane (PM) patches on a bio-functionalized porous anodic alumina (PAA) substrate. When exposed to visible light, the PM proton pumps produce a unidirectional flow of ions through the structure’s nano-pores and alter the pH of the working solution in ...

  17. Infrared spectroscopic study of photoreceptor membrane and purple membrane. Protein secondary structure and hydrogen deuterium exchange

    International Nuclear Information System (INIS)

    Infrared spectroscopy in the interval from 1800 to 1300 cm-1 has been used to investigate the secondary structure and the hydrogen/deuterium exchange behavior of bacteriorhodopsin and bovine rhodopsin in their respective native membranes. The amide I' and amide II' regions from spectra of membrane suspensions in D2O were decomposed into constituent bands by use of a curve-fitting procedure. The amide I' bands could be fit with a minimum of three theoretical components having peak positions at 1664, 1638, and 1625 cm-1 for bacteriorhodopsin and 1657, 1639, and 1625 cm-1 for rhodopsin. For both of these membrane proteins, the amide I' spectrum suggests that alpha-helix is the predominant form of peptide chain secondary structure, but that a substantial amount of beta-sheet conformation is present as well. The shape of the amide I' band was pH-sensitive for photoreceptor membranes, but not for purple membrane, indicating that membrane-bound rhodopsin undergoes a conformation change at acidic pH. Peptide hydrogen exchange of bacteriorhodopsin and rhodopsin was monitored by observing the change in the ratio of integrated absorbance (Aamide II'/Aamide I') during the interval from 1.5 to 25 h after membranes were introduced into buffered D2O. The fraction of peptide groups in a very slowly exchanging secondary structure was estimated to be 0.71 for bacteriorhodopsin at pD 7. The corresponding fraction in vertebrate rhodopsin was estimated to be less than or equal to 0.60. These findings are discussed in relationship to previous studies of hydrogen exchange behavior and to structural models for both proteins

  18. Functions of a new photoreceptor membrane. [energy conversion via halobacteria rhodopsin changes

    Science.gov (United States)

    Oesterhelt, D.; Stoeckenius, W.

    1973-01-01

    In the investigation of light responses on halobacteria phototaxis; ATP synthesis; and changes in O2 consumption, purple membrane biosynthesis, and proton translocation were found. The last three effects are discussed, which suggest that the purple membrane may function as an energy-coupling membrane for light. It is also suggested that purple membrane, through cyclic light-induced conformational changes of its bacteriorhodopsin, directly converts absorbed light energy into a proton gradient and presumably also an electric potential difference across the membrane analogous to observations in other prokaryotic cells, mitochondria, and chloroplasts.

  19. Role of Arg-72 of pharaonis Phoborhodopsin (Sensory Rhodopsin II) on its Photochemistry

    OpenAIRE

    Ikeura, Yukako; Shimono, Kazumi; Iwamoto, Masayuki; Sudo, Yuki; Kamo, Naoki

    2004-01-01

    Pharaonis phoborhodopsin (ppR, or pharaonis sensory rhodopsin II, NpsRII) is a sensor for the negative phototaxis of Natronomonas (Natronobacterium) pharaonis. Arginine 72 of ppR corresponds to Arg-82 of bacteriorhodopsin, which is a highly conserved residue among microbial rhodopsins. Using various Arg-72 ppR mutants, we obtained the following results: 1), Arg-72ppR together possibly with Asp-193 influenced the pKa of the counterion of the protonated Schiff base. 2), The M-rise became approx...

  20. FTIR spectroscopy of the M photointermediate in pharaonis rhoborhodopsin.

    OpenAIRE

    Furutani, Yuji; Iwamoto, Masayuki; Shimono, Kazumi; Kamo, Naoki; Kandori, Hideki

    2002-01-01

    pharaonis phoborhodopsin (ppR; also called pharaonis sensory rhodopsin II, psR-II) is a photoreceptor for negative phototaxis in Natronobacterium pharaonis. During the photocycle of ppR, the Schiff base of the retinal chromophore is deprotonated upon formation of the M intermediate (ppR(M)). The present FTIR spectroscopy of ppR(M) revealed that the Schiff base proton is transferred to Asp-75, which corresponds to Asp-85 in a light-driven proton-pump bacteriorhodopsin (BR). In addition, the C=...

  1. The Lifetimes of Pharaonis Phoborhodopsin Signaling States Depend on the Rates of Proton Transfers—Effects of Hydrostatic Pressure and Stopped Flow Experiments†

    OpenAIRE

    Kikukawa, Takashi; Saha, Chabita K.; Balashov, Sergei P.; Imasheva, Eleonora S.; Zaslavsky, Dmitry; Gennis, Robert B.; ABE, Takayuki; Kamo, Naoki

    2008-01-01

    Pharaonis phoborhodopsin (ppR), a negative phototaxis receptor of Natronomonas pharaonis, undergoes photocycle similar to the light-driven proton pump bacteriorhodopsin (BR), but the turnover rate is much slower due to much longer lifetimes of the M and O intermediates. The M decay was shown to become as fast as it is in BR in the L40T/F86D mutant. We examined the effects of hydrostatic pressure on the decay of these intermediates. For BR, pressure decelerated M decay but slightly affected O ...

  2. In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures

    DEFF Research Database (Denmark)

    Kjellström, Sven; Jensen, Ole Nørregaard

    2003-01-01

    A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic and...... matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS enabled the detection of the hydrophobic membrane protein bacteriorhodopsin as a component in a simple protein mixture. Peptide mixtures containing...

  3. Biosynthesis within a bubble architecture

    Science.gov (United States)

    Choi, Hyo-Jick; Montemagno, Carlo D.

    2006-05-01

    Sub-cellular compartmentalization is critical to life; it minimizes diffusion effects and enables locally high concentrations of biochemicals for improved reaction kinetics. We demonstrate an example of in vitro biochemical synthesis inside the water channels of foam using engineered artificial organelles (bacteriorhodopsin and F0F1-ATP synthase reconstituted polymer vesicles) as functional units to produce ATP. These results show that the interstitial space of bubbles serves as a metaphor for sub-cellular structure, providing a new platform for both investigating cellular metabolism and the engineering of biofunctional materials and systems.

  4. Time-resolved nonlinear polarization spectroscopy for measuring transient absorption and refraction in isotropic materials

    Science.gov (United States)

    Taranenko, Victor B.; Bazhenov, Vladimir Y.; Kulikovskaya, Olga A.

    1995-11-01

    A novel time-resolved nonlinear spectroscopic technique is described, which is based on stroboscopic registration of optical polarization transformation taking place at a vector incoherent two-wave mixing interaction in a modified Mach-Zehnder interferometer. It allows an accurate measuring of the dynamics of excitation and relaxation for real and imaginary parts of complex nonlinearity tensor components. The technique is demonstrated for measuring the light-induced change of transient absorption (delta) (alpha) e(t), (delta) (alpha) o(t) and refraction (delta) ne(t), (delta) no(t) for bacteriorhodopsin- based film pumped by linearly polarized laser pulses.

  5. One-Photon and Two-Photon Pump-Probe Spectroscopy of Photoactive Yellow Protein

    Science.gov (United States)

    Lyngnes, O.; Gibbs, H. M.; Li, C. F.; Devanathan, S. B.; Meyer, T. E.; Tollin, G.; Cusanovich, M. A.

    We present the results of nonlinear optical pump-probe experiments on photoactive yellow protein (PYP). We are able to completely bleach the 446 nm absorption peak of PYP by one-photon excitation using a cw argon laser. We calculate the corresponding index change and find it to be similar to that of bacteriorhodopsin. We also determine an upper limit to the two photon absorption cross-section of PYP by looking for bleaching of the 446 nm absorption peak under irradiation by femtosecond pulses at 820-910 nm. No TPA signal is observed.

  6. Bioferroelectricity and optical properties of biological systems

    Science.gov (United States)

    Bystrov, Vladimir; Bystrova, Natalia

    2003-08-01

    A bioferroelectric approach to analysis of ferroelectric behavior of biological systems is presented. The optical properties of nerve fibers, biomembrane ion channels, and purple membrane films containing bacteriorhodopsin are analyzed. The features, influence of the proton subsystem and proton transfer on the hydrogen-bonded biomolecular structures are analyzed within the ferroelectric liquid-crystal model and possible biomedical applications discussed. The ferroelectric behavior of biological systems and the set of various bioferroelectric effects are considered within the limits of phenomenological theory of ferroelectrics. The nonlinear response to weak actions under conditions critical to human organism is one of specific features characterizing biological objects on molecular, cell and organism levels.

  7. Moving spatial solitons in active nonlinear-optical resonators

    Science.gov (United States)

    Staliunas, K.; Taranenko, V. B.; Slekys, G.; Viselga, R.; Weiss, C. O.

    1998-01-01

    We investigate spatial solitons in a resonator with a narrow-band gain element and a saturable absorber placed in Fourier-conjugated resonator planes. Solitons are stationary or move at discrete velocities depending on the resonator tuning. The modulus of the velocity of moving solitons is fixed, but the direction of their motion is arbitrary. Solitons compete in velocity space. The experiments are conducted on a photorefractive oscillator with bacteriorhodopsin saturable absorber. Observations agree well with solutions of a general order parameter equation for such resonators.

  8. Nonlinear optical signal processing on multiwavelength sensitive materials.

    Science.gov (United States)

    Azimipour, Mehdi; Pashaie, Ramin

    2013-11-01

    Exploiting salient features in the photodynamics of specific types of light sensitive materials, a new approach is presented for realization of parallel nonlinear operations with optics. We briefly review the quantum structure and mathematical models offered for the photodynamics of two multiwavelength sensitive materials, doped crystals of lithium niobate and thick layers of bacteriorhodopsin. Next, a special mode of these dynamics in each material is investigated and a graphical design procedure is offered to produce highly nonlinear optical responses that can be dynamically reshaped via applying minimum changes in the optical setup. PMID:24177084

  9. The effects of heavy water in the proteorhodopsin photocycle

    International Nuclear Information System (INIS)

    The proton transporting photocycle of the proteorhodopsin at its normal pH (9.5) shows a marked deuterium effect. It was shown earlier that the intermediates N and PR' are responsible for the proton uptake and release. By proton-deuteron exchange the M2-N and N-PR' transitions become 2-3 times slower. On the contrary, the early μs domain is less affected than the decay part of the photocycle. The effects measured on proteorhodopsin are very similar to those measured on bacteriorhodopsin. (authors)

  10. Holographic Properties of BR-D96N Film and Its Application in Hologram Aberration Correction

    Institute of Scientific and Technical Information of China (English)

    ZHENG Yuan(郑媛); YAO Bao-Li(姚保利); WANG Ying-Li(王英利); MENKE Neimule(门克内木乐); LEI Ming(雷铭); CHEN Guo-Fu(陈国夫); Norbert HAMPP

    2003-01-01

    A biophotochromic material, i.e., genetic mutant bacteriorhodopsin (BR-D96N), was experimentally studied on its holographic recording properties. The saturation absorption curve and the diffraction efficiency curve were measured respectively. As holographic storage application, reflection type polarization holograms were recorded on the BR-D96N film. The 173° configuration between the object and reference beams proves that the spatial resolution of the film is over 6000 lines/mm. By using phase conjugate wave of reference beam as reconstruction beam, the distorted object image introduced by the optical components and the defects in the recording medium can be well corrected.

  11. [Biomimetic sensors in biomedical research].

    Science.gov (United States)

    Gayet, Landry; Lenormand, Jean-Luc

    2015-01-01

    The recent research on both the synthesis of membrane proteins by cell-free systems and the reconstruction of planar lipid membranes, has led to the development of a cross-technology to produce biosensors or filters. Numerous biomimetic membranes are currently being standardized and used by the industry, such as filters containing aquaporin for water desalination, or used in routine at the laboratory scale, for example the bacteriorhodopsin as a light sensor. In the medical area, several fields of application of these biomimetic membranes are under consideration today, particularly for the screening of therapeutic molecules and for the developing of new tools in diagnosis, patient monitoring and personalized medicine. PMID:26152170

  12. Proton Pumps: Mechanism of Action and Applications

    Science.gov (United States)

    Lanyi, Janos K.; Pohorille, Andrew; DeVincenzi, Donald L. (Technical Monitor)

    2001-01-01

    Recent progress in understanding molecular structures and mechanisms of action of proton pumps has paved the way to their novel applications in biotechnology. Proton pumps, in particular bacteriorhodopsin and ATP synthases, are capable of continuous, renewable conversion of light to chemical, mechanical or electrical energy, which can be used in macro- or nano-scale devices. The capability of protein systems incorporated into liposomes to generate ATP, which can be further used to drive chemical reactions, and to act as molecular motors has been already demonstrated. Other possible applications of such biochemical devices include targeted drug delivery and biocatalytic re actors. All these devices might prove superior to their inorganic alternatives.

  13. Crystallographic Structure of Xanthorhodopsin, the Light-Driven Proton Pump With a Dual Chromophore

    Energy Technology Data Exchange (ETDEWEB)

    Luecke, H.; Schobert, B.; Stagno, J.; Imasheva, E.S.; Wang, J.M.; Balashov, S.P.; Lanyi, J.K.

    2009-05-19

    Homologous to bacteriorhodopsin and even more to proteorhodopsin, xanthorhodopsin is a light-driven proton pump that, in addition to retinal, contains a noncovalently bound carotenoid with a function of a light-harvesting antenna. We determined the structure of this eubacterial membrane protein-carotenoid complex by X-ray diffraction, to 1.9-{angstrom} resolution. Although it contains 7 transmembrane helices like bacteriorhodopsin and archaerhodopsin, the structure of xanthorhodopsin is considerably different from the 2 archaeal proteins. The crystallographic model for this rhodopsin introduces structural motifs for proton transfer during the reaction cycle, particularly for proton release, that are dramatically different from those in other retinal-based transmembrane pumps. Further, it contains a histidine-aspartate complex for regulating the pK{sub a} of the primary proton acceptor not present in archaeal pumps but apparently conserved in eubacterial pumps. In addition to aiding elucidation of a more general proton transfer mechanism for light-driven energy transducers, the structure defines also the geometry of the carotenoid and the retinal. The close approach of the 2 polyenes at their ring ends explains why the efficiency of the excited-state energy transfer is as high as {approx}45%, and the 46{sup o} angle between them suggests that the chromophore location is a compromise between optimal capture of light of all polarization angles and excited-state energy transfer.

  14. Photoreversible conformational changes in membrane proteins using light-responsive surfactants.

    Science.gov (United States)

    Zhang, Jing; Wang, Shao-Chun; Lee, C Ted

    2009-06-25

    Photoreversible control of the conformation of bacteriorhodopsin in the presence of a light-responsive surfactant is demonstrated through combined UV-vis, FT-IR, and (31)P NMR spectroscopy and dynamic light scattering (DLS) measurements. The azobenzene-based surfactant photoisomerizes upon 434 nm visible (trans, relatively hydrophobic) and 350 nm UV (cis, relatively hydrophilic) illumination, allowing surfactant micellization to be reversibly controlled. This leads to partitioning of the membrane protein into micelles in the unfolded state under visible light, while UV light leads to solubilization of the protein within purple membrane bilayers in the folded state. A three-stage model of purple membrane-photosurfactant interactions is examined through NMR and DLS measurements. Phototriggered unfolding of bacteriorhodopsin, occurring through alpha(II) --> alpha(I) and reverse beta-turn --> extended beta-strand transitions, requires approximately 20 s for completion, while light-induced refolding requires a somewhat longer 80 s as the membrane protein repartitions into the reformed bilayer membrane. Each of these conformational changes can be precisely and reversibly controlled with simple light illumination, providing a novel technique to probe membrane protein folding. PMID:19485396

  15. Metabolism, Physiology and Biotechnological Applications of Halobacteria

    Directory of Open Access Journals (Sweden)

    Oleg Mosin

    2015-06-01

    Full Text Available Halophiles (lat. “salt-loving” is the taxonomic group of extreme aerobic obligate Gram-negative microorganisms that live in conditions of high salinity – in the seas, salt lakes, saline soils etc. These microorganisms are known to reddish patina on products, preserved with using large quantities of salt (NaCl. Halophiles were isolated for the first time at the beginning of the XX century from the marine flora estuary mud, but their systematic study was started only at the end of the second decade of the XX century. The internal environment of the human body is not suitable for existence of halobacteria, since none of them are known to have pathogenic forms. Halobacteria have great practical potential for using in molecular bioelectronics and bio-nanotechnology due to their unique ability to convert the energy of sunlight into electrochemical energy of protons H+ due to the presence in their cells a special photo transforming retinal containing integral protein – bacteriorhodopsin, the mechanism of action of which has been currently studied in detail. This article describes the characteristics of the metabolism and physiology of halophilic bacteria, as well as a method of biosynthesis and preparation of bacteriorhodopsin from purple membranes of cells of the extreme photoorganotrophic halobacterium Halobacterium halobium.

  16. Specificity of anion-binding in the substrate-pocket ofbacteriorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Facciotti, Marc T.; Cheung, Vincent S.; Lunde, Christopher S.; Rouhani, Shahab; Baliga, Nitin S.; Glaeser, Robert M.

    2003-08-30

    The structure of the D85S mutant of bacteriorhodopsin with a nitrate anion bound in the Schiff-base binding site, and the structure of the anion-free protein have been obtained in the same crystal form. Together with the previously solved structures of this anion pump, in both the anion-free state and bromide-bound state, these new structures provide insight into how this mutant of bacteriorhodopsin is able to bind a variety of different anions in the same binding pocket. The structural analysis reveals that the main structural change that accommodates different anions is the repositioning of the polar side-chain of S85. On the basis of these x-ray crystal structures, the prediction is then made that the D85S/D212N double mutant might bind similar anions and do so over a broader pH range than does the single mutant. Experimental comparison of the dissociation constants, K{sub d}, for a variety of anions confirms this prediction and demonstrates, in addition, that the binding affinity is dramatically improved by the D212N substitution.

  17. Crystallographic Structure of Xanthorhodopsin, the Light-Driven Proton Pump With a Dual Chromophore

    International Nuclear Information System (INIS)

    Homologous to bacteriorhodopsin and even more to proteorhodopsin, xanthorhodopsin is a light-driven proton pump that, in addition to retinal, contains a noncovalently bound carotenoid with a function of a light-harvesting antenna. We determined the structure of this eubacterial membrane protein-carotenoid complex by X-ray diffraction, to 1.9-(angstrom) resolution. Although it contains 7 transmembrane helices like bacteriorhodopsin and archaerhodopsin, the structure of xanthorhodopsin is considerably different from the 2 archaeal proteins. The crystallographic model for this rhodopsin introduces structural motifs for proton transfer during the reaction cycle, particularly for proton release, that are dramatically different from those in other retinal-based transmembrane pumps. Further, it contains a histidine-aspartate complex for regulating the pKa of the primary proton acceptor not present in archaeal pumps but apparently conserved in eubacterial pumps. In addition to aiding elucidation of a more general proton transfer mechanism for light-driven energy transducers, the structure defines also the geometry of the carotenoid and the retinal. The close approach of the 2 polyenes at their ring ends explains why the efficiency of the excited-state energy transfer is as high as ∼45%, and the 46o angle between them suggests that the chromophore location is a compromise between optimal capture of light of all polarization angles and excited-state energy transfer

  18. Slow alpha helix formation during folding of a membrane protein.

    Science.gov (United States)

    Riley, M L; Wallace, B A; Flitsch, S L; Booth, P J

    1997-01-01

    Very little is known about the folding of proteins within biological membranes. A "two-stage" model has been proposed on thermodynamic grounds for the folding of alpha helical, integral membrane proteins, the first stage of which involves formation of transmembrane alpha helices that are proposed to behave as autonomous folding domains. Here, we investigate alpha helix formation in bacteriorhodopsin and present a time-resolved circular dichroism study of the slow in vitro folding of this protein. We show that, although some of the protein's alpha helices form early, a significant part of the protein's secondary structure appears to form late in the folding process. Over 30 amino acids, equivalent to at least one of bacteriorhodopsin's seven transmembrane segments, slowly fold from disordered structures to alpha helices with an apparent rate constant of about 0.012 s-1 at pH 6 or 0.0077 s-1 at pH 8. This is a rate-limiting step in protein folding, which is dependent on the pH and the composition of the lipid bilayer. PMID:8993333

  19. Protein-Based Three-Dimensional Memories and Associative Processors

    Science.gov (United States)

    Birge, Robert

    2008-03-01

    The field of bioelectronics has benefited from the fact that nature has often solved problems of a similar nature to those which must be solved to create molecular electronic or photonic devices that operate with efficiency and reliability. Retinal proteins show great promise in bioelectronic devices because they operate with high efficiency (˜0.65%), high cyclicity (>10^7), operate over an extended wavelength range (360 -- 630 nm) and can convert light into changes in voltage, pH, absorption or refractive index. This talk will focus on a retinal protein called bacteriorhodopsin, the proton pump of the organism Halobacterium salinarum. Two memories based on this protein will be described. The first is an optical three-dimensional memory. This memory stores information using volume elements (voxels), and provides as much as a thousand-fold improvement in effective capacity over current technology. A unique branching reaction of a variant of bacteriorhodopsin is used to turn each protein into an optically addressed latched AND gate. Although three working prototypes have been developed, a number of cost/performance and architectural issues must be resolved prior to commercialization. The major issue is that the native protein provides a very inefficient branching reaction. Genetic engineering has improved performance by nearly 500-fold, but a further order of magnitude improvement is needed. Protein-based holographic associative memories will also be discussed. The human brain stores and retrieves information via association, and human intelligence is intimately connected to the nature and enormous capacity of this associative search and retrieval process. To a first order approximation, creativity can be viewed as the association of two seemingly disparate concepts to form a totally new construct. Thus, artificial intelligence requires large scale associative memories. Current computer hardware does not provide an optimal environment for creating artificial

  20. Controlled in meso phase crystallization--a method for the structural investigation of membrane proteins.

    Directory of Open Access Journals (Sweden)

    Jan Kubicek

    Full Text Available We investigated in meso crystallization of membrane proteins to develop a fast screening technology which combines features of the well established classical vapor diffusion experiment with the batch meso phase crystallization, but without premixing of protein and monoolein. It inherits the advantages of both methods, namely (i the stabilization of membrane proteins in the meso phase, (ii the control of hydration level and additive concentration by vapor diffusion. The new technology (iii significantly simplifies in meso crystallization experiments and allows the use of standard liquid handling robots suitable for 96 well formats. CIMP crystallization furthermore allows (iv direct monitoring of phase transformation and crystallization events. Bacteriorhodopsin (BR crystals of high quality and diffraction up to 1.3 Å resolution have been obtained in this approach. CIMP and the developed consumables and protocols have been successfully applied to obtain crystals of sensory rhodopsin II (SRII from Halobacterium salinarum for the first time.

  1. The rhodopsin-like pigments of halobacteria - Light-energy and signal transducers in an archaebacterium

    Science.gov (United States)

    Stoeckenius, W.

    1985-01-01

    Three, small retinylidene proteins observed in halobacteria are described. The characteristics of bacteriorhodopsin (bR), which is synthesized during low O2 tension and intense illumination, and the role of bR in the cyclic photoreactions that translocate protons are examined. The detected light-driven chloride influx pigment, halorhodopsin (hR), is also capable of light-driven ion translocation; the hR transport reactions which are chloride dependent and involve isomerization are studied. The sensory photosystem of halobacteria and the receptor functions of the retinal pigment slow rhodopsin are discussed. The similarity of the choromphore structure and photoreactions, and the evolutionary relation between halobacteria and animal pigments are considered.

  2. A Photoisomerizing Rhodopsin Mimic Observed at Atomic Resolution.

    Science.gov (United States)

    Nosrati, Meisam; Berbasova, Tetyana; Vasileiou, Chrysoula; Borhan, Babak; Geiger, James H

    2016-07-20

    The members of the rhodopsin family of proteins are involved in many essential light-dependent processes in biology. Specific photoisomerization of the protein-bound retinylidene PSB at a specified wavelength range of light is at the heart of all of these systems. Nonetheless, it has been difficult to reproduce in an engineered system. We have developed rhodopsin mimics, using intracellular lipid binding protein family members as scaffolds, to study fundamental aspects of protein/chromophore interactions. Herein we describe a system that specifically isomerizes the retinylidene protonated Schiff base both thermally and photochemically. This isomerization has been characterized at atomic resolution by quantitatively interconverting the isomers in the crystal both thermally and photochemically. This event is accompanied by a large pKa change of the imine similar to the pKa changes observed in bacteriorhodopsin and visual opsins during isomerization. PMID:27310917

  3. Investigation of the chromophore binding cavity in the 11-cis acceptable microbial rhodopsin MR

    Science.gov (United States)

    Mori, Arisa; Yagasaki, Jin; Homma, Michio; Reissig, Louisa; Sudo, Yuki

    2013-06-01

    Rhodopsins are photoactive molecules functioning as photo-energy or photo-signal converters with the chromophore retinal. Recently we characterized a unique microbial rhodopsin (middle rhodopsin, MR) which can also bind 11-cis retinal besides all-trans and 13-cis retinal at a particular ratio. In this study, we investigated the structural characteristics around the retinal binding cavity in MR. The results suggest that the space of the retinal binding site of MR is less restricted to the retinal chromophore and the presence of the 11-cis conformer is regulated by the residues located around the retinal. Furthermore, although the triple mutant of MR has identical residues with the well-studied microbial rhodopsin bacteriorhodopsin (BR) within 5 Å from the retinal, the absorption maximum and retinal composition of MR did not reach those of BR, indicating that some long-range effect(s) (>5 Å) is also important for the maintenance of the chemical properties of MR.

  4. Chapter IV: ultrafast biochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Chergui, M. [Swiss Federal Institute of Technology (EPFL), Lausanne (Switzerland); Kjelstrup, S. [Norwegian University of Science and Technology (NTNU), Trondheim (Norway); Meuwly, M. [Universitaet Basel, Basel (Switzerland); Schuler, B. [University of Zuerich (ETH), Zurich (Switzerland); Thor, J. van [Imperial College London (IC), London (United Kingdom)

    2009-09-15

    The whole report issued by the Paul Scherrer Institute (PSI) in Switzerland takes a look at the scientific opportunities offered by the institute's SwissFEL X-ray Laser facility. In this sixth part, initial events and fluctuations in biochemical processes at the atomic scale are discussed. Sub-nanosecond processes are fundamental to biochemistry and will be accessible to the ultra-short pulses of the SwissFEL. Time and length scales of biochemical reactions are discussed, as is the photo-initiation of biochemical processes. Time-resolved measurement techniques are looked at. Fluorescence resonant energy transfer is discussed. As an example, the photo cycle of bacteriorhodopsin is examined. The dynamics of protein folding and catalytic action are also looked at. Mesoscopic non-equilibrium thermodynamics is discussed

  5. Probing how initial retinal configuration controls photochemical dynamics in retinal proteins

    Directory of Open Access Journals (Sweden)

    Sheves M.

    2013-03-01

    Full Text Available The effects of the initial retinal configuration and the active isomerization coordinate on the photochemistry of retinal proteins (RPs are assessed by comparing photochemical dynamics of two stable retinal ground state configurations (all-trans,15-anti vs. 13-cis,15-syn, within two RPs: Bacteriorhodopsin (BR and Anabaena Sensory Rhodopsin (ASR. Hyperspectral pump-probe spectroscopy shows that photochemistry starting from 13-cis retinal in both proteins is 3-10 times faster than when started in the all-trans state, suggesting that the hastening is ubiquitous to microbial RPs, regardless of their different biological functions and origin. This may also relate to the known disparity of photochemical rates between microbial RPs and visual pigments. Importance and possible underlying mechanisms are discussed as well.

  6. Photoreceptors for a light biotransducer: a comparative study of the electrical responses of two (type-1)-opsins

    CERN Document Server

    Alfinito, E; Reggiani, L; Lee, K

    2013-01-01

    The increasing interest in photoactivated proteins as natural replacement of standard inorganic materials in photocells drives to the compared analysis of bacteriorhodopsin and proteorhodopsin, two widely diffused proteins belonging to the family of \\textit{type-1} opsins. These proteins share similar behaviours but exhibit relevant differences in the sequential chain of the amino acids constituting their tertiary structure. The use of an impedance network analogue to model the protein main features provides a microscopic interpretation of a set of experiments on their photoconductance properties. In particular, this model links the protein electrical responses to the tertiary structure and to the interactions among neighbouring amino acids. The same model is also used to predict the small-signal response in terms of the Nyquist plot. Interesting enough, these rhodopsins are found to behave like a wide gap semiconductor with intrinsic conductivities of the order of $10^{-7}$ S/cm.

  7. Steady state emission of the fluorescent intermediate of Anabaena Sensory Rhodopsin as a function of light adaptation conditions

    Science.gov (United States)

    Cheminal, A.; Léonard, J.; Kim, S. Y.; Jung, K.-H.; Kandori, H.; Haacke, S.

    2013-11-01

    Steady-state fluorescence measurements of the first excited state of the anabaena sensory rhodopsin (ASR), and Bacteriorhodopsin are reported for different light stabilization conditions, including the dark-adapted state. We determine the fluorescence spectra of both all-trans (AT), and 13-cis (13C) protonated Schiff base of retinal, and compare the effect of the proteins. Referenced against the fluorescence quantum yield of AT-bR (2.5 × 10-4) we find for AT-ASR, 13C-ASR, and 13C-bR the values of 3.3 × 10-4, 0.8 × 10-4, and 1.7 × 10-4, respectively. Using reported excited state lifetimes, the radiative rates are deduced, and their differences discussed on the basis of a configuration-dependent oscillator strength.

  8. Current-voltage characteristics of seven-helix proteins from a cubic array of amino acids

    Science.gov (United States)

    Alfinito, Eleonora; Reggiani, Lino

    2016-06-01

    The electrical properties of a set of seven-helix transmembrane proteins, whose space arrangement [three-dimensional (3D) structure] is known, are investigated by using regular arrays of the amino acids. These structures, specifically cubes, have topological features similar to those shown by the chosen proteins. The theoretical results show a good agreement between the predicted current-voltage characteristics obtained from a cubic array and those obtained from a detailed 3D structure. The agreement is confirmed by available experiments on bacteriorhodopsin. Furthermore, all the analyzed proteins are found to share the same critical behavior of the voltage-dependent conductance and of its variance. In particular, the cubic arrangement evidences a short plateau of the excess conductance and its variance at high voltages. The results of the present investigation show the possibility to predict the I -V characteristics of a multiple-protein sample even in the absence of detailed knowledge of the proteins' 3D structure.

  9. Characterization and isolation of a light driven sodium pump from membranes of Halobacterium halobium. Final technical progress report

    International Nuclear Information System (INIS)

    We investigated three aspects of the light driven sodium pump (halorhodopsin, which appear to be crucial to our understanding of the mechanisms employed by Halobacterium halobium and to further investigate this unique system of energy conservation. We characterized the molecular mechanisms of transmembrane sodium transport in vesicles from H. halobium with particular reference to the mechanism of couplins of light energy to net sodium translocation. We develop procedures and techniques for extracting the components of the light driven sodium pump from membranes and incorporating them into artificial membrane systems. We examine the mechanism of conversion of bacteriorhodopsin from an active to an inactive form in membrane vesicles and to relate this alternative state of this pigment to the presence of the light driven sodium pump

  10. Partial purification of the chloroplast ATP synthase from Chlamydomonas reinhardtii and the cloning and sequencing of a cDNA encoding the gamma subunit

    International Nuclear Information System (INIS)

    The chloroplast ATP synthase was partially purified from the green alga Chlamydomonas reinhardtii by extracting membranes with deoxycholate and KCl, followed by centrifugation and ammonium sulfate fractionation of the supernatant. The enzyme assay involved the reconstitution of such fractions with bacteriorhodopsin and soybean phospholipids to form vesicles capable of light-dependent [32P]-phosphate esterification. A cDNA for the gamma subunit from Chlamydomonas was isolated, expressed in vitro and sequenced. It contains the entire coding region for the gamma subunit precursor. A 35 amino acid long transit peptide resides at the NH2-terminus of a 323 amino acid long mature peptide that is 77% similar to the spinach gamma subunit. Six cysteines were found; three were conserved in Chlamydomonas and spinach

  11. High-Speed Atomic Force Microscopy

    Science.gov (United States)

    Ando, Toshio; Uchihashi, Takayuki; Kodera, Noriyuki

    2012-08-01

    The technology of high-speed atomic force microscopy (HS-AFM) has reached maturity. HS-AFM enables us to directly visualize the structure and dynamics of biological molecules in physiological solutions at subsecond to sub-100 ms temporal resolution. By this microscopy, dynamically acting molecules such as myosin V walking on an actin filament and bacteriorhodopsin in response to light are successfully visualized. High-resolution molecular movies reveal the dynamic behavior of molecules in action in great detail. Inferences no longer have to be made from static snapshots of molecular structures and from the dynamic behavior of optical markers attached to biomolecules. In this review, we first describe theoretical considerations for the highest possible imaging rate, then summarize techniques involved in HS-AFM and highlight recent imaging studies. Finally, we briefly discuss future challenges to explore.

  12. Determination of deuterium-labeled tryptophan in proteins by means of high-performance liquid chromatography and thermospray mass spectrometry

    International Nuclear Information System (INIS)

    In order to develop direct methods for determining the extent of metabolic incorporation of isotopically labeled amino acids into a protein, the determination of deuterated tryptophan in [2H5]tryptophan-bacteriorhodopsin was investigated. The isotopically modified protein was subjected to alkaline hydrolysis. After phenyl isothiocyanate derivatization of the hydrolysate, the mixture was separated by reversed-phase liquid chromatography. Field desorption mass spectrometry and thermospray mass spectrometry were investigated for their ability to determine the ratio between [2H5]tryptophan and total tryptophan in the collected fractions. In order to check the procedure a set of known tryptophan/[2H5]tryptophan mixtures were passed through the same derivatization, HPLC separation, and lyophilization procedure as used for the biological samples

  13. {sup 13}C- and {sup 1}H-detection under fast MAS for the study of poorly available proteins: application to sub-milligram quantities of a 7 trans-membrane protein

    Energy Technology Data Exchange (ETDEWEB)

    Dannatt, Hugh R. W.; Taylor, Garrick F.; Varga, Krisztina; Higman, Victoria A.; Pfeil, Marc-Philipp; Asilmovska, Lubica; Judge, Peter J.; Watts, Anthony, E-mail: anthony.watts@bioch.ox.ac.uk [University of Oxford, Department of Biochemistry (United Kingdom)

    2015-05-15

    We demonstrate that {sup 13}C-detected spectra recorded using fast (60 kHz) magic angle spinning on sub-milligram (<10 μmol) quantities of a protonated 7 trans-membrane helix protein (bacteriorhodopsin) in its native lipid environment are comparable in sensitivity and resolution to those recorded using 15-fold larger sample volumes with conventional solid state NMR methodology. We demonstrate the utility of proton-detected measurements which yield narrow {sup 1}H linewidths under these conditions, and that no structural alterations are observed. We propose that these methods will prove useful to gain structural information on membrane proteins with poor availability, which can be studied in their native lipid environments.

  14. Raman-Nath thin gratings of low-saturated dynamic recording materials

    Directory of Open Access Journals (Sweden)

    S.Bugaychuk

    2002-03-01

    Full Text Available The holographic kinetics of organic polymer thin films with low-saturated absorption are considered both for self-diffraction and diffraction of a weak probe beam in Raman-Nath conditions. The coupling effect between different diffracted orders is taking into account. The coupling effect results in considerable increase of the diffraction efficiency of self-diffracted beams compared with the diffraction of the probe beam were shown. For the region of deep saturation the maximum diffraction efficiency is higher than in the region far from the saturation. The maximum of diffraction efficiency is reached for considerable shorter time in grating recording kinetics in the region of deep saturation. This opens the perspective of using those materials for novelty filter. The experimental results on low-saturated dry bacteriorhodopsin films well agree the theoretical predictions.

  15. A network model to investigate structural and electrical properties of proteins

    CERN Document Server

    Alfinito, E; Reggiani, L

    2007-01-01

    One of the main trend in to date research and development is the miniaturization of electronic devices. In this perspective, integrated nanodevices based on proteins or biomolecules are attracting a major interest. In fact, it has been shown that proteins like bacteriorhodopsin and azurin, manifest electrical properties which are promising for the development of active components in the field of molecular electronics. Here we focus on two relevant kinds of proteins: The bovine rhodopsin, prototype of GPCR protein, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer disease. Both these proteins exert their functioning starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different electrical response associated with the diff...

  16. A 250 GHz gyrotron with a 3 GHz tuning bandwidth for dynamic nuclear polarization.

    Science.gov (United States)

    Barnes, Alexander B; Nanni, Emilio A; Herzfeld, Judith; Griffin, Robert G; Temkin, Richard J

    2012-08-01

    We describe the design and implementation of a novel tunable 250 GHz gyrotron oscillator with >10 W output power over most of a 3 GHz band and >35 W peak power. The tuning bandwidth and power are sufficient to generate a >1 MHz nutation frequency across the entire nitroxide EPR lineshape for cross effect DNP, as well as to excite solid effect transitions utilizing other radicals, without the need for sweeping the NMR magnetic field. Substantially improved tunability is achieved by implementing a long (23 mm) interaction cavity that can excite higher order axial modes by changing either the magnetic field of the gyrotron or the cathode potential. This interaction cavity excites the rotating TE(₅,₂,q) mode, and an internal mode converter outputs a high-quality microwave beam with >94% Gaussian content. The gyrotron was integrated into a DNP spectrometer, resulting in a measured DNP enhancement of 54 on the membrane protein bacteriorhodopsin. PMID:22743211

  17. A 250 GHz gyrotron with a 3 GHz tuning bandwidth for dynamic nuclear polarization

    Science.gov (United States)

    Barnes, Alexander B.; Nanni, Emilio A.; Herzfeld, Judith; Griffin, Robert G.; Temkin, Richard J.

    2012-08-01

    We describe the design and implementation of a novel tunable 250 GHz gyrotron oscillator with >10 W output power over most of a 3 GHz band and >35 W peak power. The tuning bandwidth and power are sufficient to generate a >1 MHz nutation frequency across the entire nitroxide EPR lineshape for cross effect DNP, as well as to excite solid effect transitions utilizing other radicals, without the need for sweeping the NMR magnetic field. Substantially improved tunability is achieved by implementing a long (23 mm) interaction cavity that can excite higher order axial modes by changing either the magnetic field of the gyrotron or the cathode potential. This interaction cavity excites the rotating TE5,2,q mode, and an internal mode converter outputs a high-quality microwave beam with >94% Gaussian content. The gyrotron was integrated into a DNP spectrometer, resulting in a measured DNP enhancement of 54 on the membrane protein bacteriorhodopsin.

  18. Genetic analysis of the br gene in halophilic archaea isolated from Xinjiang region, China

    Institute of Scientific and Technical Information of China (English)

    Xiaohong XU; Min WU; Huibin ZHANG; Zhihu LIU

    2008-01-01

    Some novel members of extremely halophilic archaea, strains AJ 11, AJ 12 and AJ 13, were isolated from the Aularz Lake located in the Altun Mountain National Nature Reserve of Xinjiang, Uygur Autonomous Region in China. Partial DNA fragments encoding a bacteriorho-dopsin (BR), as well as for 16S rRNA of isolated strains, were amplified by PCR and their DNA sequences were determined subsequently. On the basis of homology and phylogenetic analysis of the 16S rDNA, we thought that the isolated strains forming a microbiological population are the members of the genus Natrinema. The results of genetic analysis, such as GC content, transition/transver-sion (Ti/Tv) rate ratios and synonymous substitution rates (Ks) indicate that the br fragments, with a high level of genetic divergence, are faced with both purifying selection and bias mutation pressure. The study provides the basis for use of species and BR proteins resources.

  19. A light-driven three-dimensional plasmonic nanosystem that translates molecular motion into reversible chiroptical function

    Science.gov (United States)

    Kuzyk, Anton; Yang, Yangyang; Duan, Xiaoyang; Stoll, Simon; Govorov, Alexander O.; Sugiyama, Hiroshi; Endo, Masayuki; Liu, Na

    2016-02-01

    Nature has developed striking light-powered proteins such as bacteriorhodopsin, which can convert light energy into conformational changes for biological functions. Such natural machines are a great source of inspiration for creation of their synthetic analogues. However, synthetic molecular machines typically operate at the nanometre scale or below. Translating controlled operation of individual molecular machines to a larger dimension, for example, to 10-100 nm, which features many practical applications, is highly important but remains challenging. Here we demonstrate a light-driven plasmonic nanosystem that can amplify the molecular motion of azobenzene through the host nanostructure and consequently translate it into reversible chiroptical function with large amplitude modulation. Light is exploited as both energy source and information probe. Our plasmonic nanosystem bears unique features of optical addressability, reversibility and modulability, which are crucial for developing all-optical molecular devices with desired functionalities.

  20. 13C- and 1H-detection under fast MAS for the study of poorly available proteins: application to sub-milligram quantities of a 7 trans-membrane protein

    International Nuclear Information System (INIS)

    We demonstrate that 13C-detected spectra recorded using fast (60 kHz) magic angle spinning on sub-milligram (<10 μmol) quantities of a protonated 7 trans-membrane helix protein (bacteriorhodopsin) in its native lipid environment are comparable in sensitivity and resolution to those recorded using 15-fold larger sample volumes with conventional solid state NMR methodology. We demonstrate the utility of proton-detected measurements which yield narrow 1H linewidths under these conditions, and that no structural alterations are observed. We propose that these methods will prove useful to gain structural information on membrane proteins with poor availability, which can be studied in their native lipid environments

  1. Rapid thermal tuning of chromophore structure in membrane protein.

    Science.gov (United States)

    Wang, Jianping; El-Sayed, Mostafa A

    2009-04-01

    We show that the configuration and the optical property of the retinal chromophore in bacteriorhodopsin (bR) can be tuned dynamically from the all-trans configuration to the 13-cis by using a nanosecond laser-induced temperature-jump. The rapid bleach in the visible absorption optical density of retinal has an apparent formation time of ca. 170 ns, whereas the relaxation process finishes within tens of ms. The dynamical transition of retinal from the all-trans to 13-cis species is believed to occur as a result of rapid protein conformational change especially in the vicinity of retinal binding site. Our study reveals the intrinsic dynamical aspect of the retinal chromophore with respect to the protein structure. PMID:19275202

  2. Laser-diode-based joint transform correlator for fingerprint identification

    Science.gov (United States)

    Lal, Amit K.; Zang, De Yu; Millerd, James E.

    1999-01-01

    A laser-diode-based joint transform correlator (JTC) is reported here for the identification and discrimination of fingerprints. The system employs compact and inexpensive laser diodes as the light sources and a bacteriorhodopsin (BR) film in the Fourier plane, which can record the joint power spectrum without the need for expensive spatial light modulators or CCD cameras. The BR film also introduces nonlinearities in the Fourier plane which can improve JTC performance. In addition, real-time, all-optical programmable spatial filtering is demonstrated to improve the discrimination of the system. We present computer modeling and experimental results of this optical correlator, which shows excellent potential for the identification and discrimination of fingerprints.

  3. H+-type and OH--type biological protonic semiconductors and complementary devices

    Science.gov (United States)

    Deng, Yingxin; Josberger, Erik; Jin, Jungho; Rousdari, Anita Fadavi; Helms, Brett A.; Zhong, Chao; Anantram, M. P.; Rolandi, Marco

    2013-10-01

    Proton conduction is essential in biological systems. Oxidative phosphorylation in mitochondria, proton pumping in bacteriorhodopsin, and uncoupling membrane potentials by the antibiotic Gramicidin are examples. In these systems, H+ hop along chains of hydrogen bonds between water molecules and hydrophilic residues - proton wires. These wires also support the transport of OH- as proton holes. Discriminating between H+ and OH- transport has been elusive. Here, H+ and OH- transport is achieved in polysaccharide- based proton wires and devices. A H+- OH- junction with rectifying behaviour and H+-type and OH--type complementary field effect transistors are demonstrated. We describe these devices with a model that relates H+ and OH- to electron and hole transport in semiconductors. In turn, the model developed for these devices may provide additional insights into proton conduction in biological systems.

  4. Protons and how they are transported by proton pumps

    DEFF Research Database (Denmark)

    Buch-Pedersen, Morten Jeppe; Pedersen, Bjørn Panyella; Nissen, Poul;

    2008-01-01

    The very high mobility of protons in aqueous solutions demands special features of membrane proton transporters to sustain efficient yet regulated proton transport across biological membranes. By the use of the chemical energy of ATP, plasma-membrane-embedded ATPases extrude protons from cells of...... plants and fungi to generate electrochemical proton gradients. The recently published crystal structure of a plasma membrane H(+)-ATPase contributes to our knowledge about the mechanism of these essential enzymes. Taking the biochemical and structural data together, we are now able to describe the basic...... molecular components that allow the plasma membrane proton H(+)-ATPase to carry out proton transport against large membrane potentials. When divergent proton pumps such as the plasma membrane H(+)-ATPase, bacteriorhodopsin, and F(O)F(1) ATP synthase are compared, unifying mechanistic premises for biological...

  5. Model Construction and Analysis of Respiration in Halobacterium salinarum.

    Directory of Open Access Journals (Sweden)

    Cherryl O Talaue

    Full Text Available The archaeon Halobacterium salinarum can produce energy using three different processes, namely photosynthesis, oxidative phosphorylation and fermentation of arginine, and is thus a model organism in bioenergetics. Compared to its bacteriorhodopsin-driven photosynthesis, less attention has been devoted to modeling its respiratory pathway. We created a system of ordinary differential equations that models its oxidative phosphorylation. The model consists of the electron transport chain, the ATP synthase, the potassium uniport and the sodium-proton antiport. By fitting the model parameters to experimental data, we show that the model can explain data on proton motive force generation, ATP production, and the charge balancing of ions between the sodium-proton antiporter and the potassium uniport. We performed sensitivity analysis of the model parameters to determine how the model will respond to perturbations in parameter values. The model and the parameters we derived provide a resource that can be used for analytical studies of the bioenergetics of H. salinarum.

  6. Creating electrochemical gradients by light: from bio-inspired concepts to photoelectric conversion.

    Science.gov (United States)

    Xie, Xiaojiang; Bakker, Eric

    2014-10-01

    Light is harvested by natural photosynthetic systems to generate electrochemical gradients that power various reactions. Implementing nature's lessons in photosynthesis holds great promise for technological advances. With a focus on designs and concepts, recent progress in generating electrochemical gradients by light, mimicking the two general types of photosynthetic centers in nature that make use of either light-induced charge separation or photo-isomerization are summarized here. Light induced electrochemical gradients pave new ways for photoelectric conversion. While extensive research in this direction has focused on light-induced charge separation, recent work has shown that energy conversion based on photo-isomerization is very promising. Photoswitchable compounds have been found in nature, such as the retinal molecule in bacteriorhodopsin. These compounds may form an attractive molecular basis for future progress in this field. PMID:25116853

  7. Reconstitution of halorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Kong, T.

    1989-11-01

    Halobacterium halobium contains a family of retinal-bound proteins: bacteriorhodopsin (bR) which mediates phototrophic growth as a light-riven proton pump, halorhodopsin (hR) which is a light-driven chloride pump, and one or more sensory rhodopsins (sR) which mediate a phototactic response. Two-dimensional crystallization of halorhodopsin has been attempted though the reconstitution of purified halorhodopsin with purple membrane lipid for electron microscopy work. The first important step for crystallization is to get a homogeneous protein which is pure and not denatured. Homogeneous halorhodopsin has been obtained by a modification of existing purification methods. Some nice looking membrane patches which have the same density as purple membrane have been obtained. But unfortunately, they are not crystalline. The procedure of hR reconstitution is described in detail and some other strategies to induce the protein crystal in the reconstituted membrane are discussed in this dissertation. 76 refs., 20 figs., 6 tabs.

  8. Dynamic Nuclear Polarization enhanced NMR at 187 GHz/284 MHz using an Extended Interaction Klystron amplifier

    Science.gov (United States)

    Kemp, Thomas F.; Dannatt, Hugh R. W.; Barrow, Nathan S.; Watts, Anthony; Brown, Steven P.; Newton, Mark E.; Dupree, Ray

    2016-04-01

    A Dynamic Nuclear Polarisation (DNP) enhanced solid-state Magic Angle Spinning (MAS) NMR spectrometer which uses a 187 GHz (corresponding to 1H NMR frequency of 284 MHz) Extended Interaction Klystron (EIK) amplifier as the microwave source is briefly described. Its performance is demonstrated for a biomolecule (bacteriorhodopsin), a pharmaceutical, and surface functionalised silica. The EIK is very compact and easily incorporated into an existing spectrometer. The bandwidth of the amplifier is sufficient that it obviates the need for a sweepable magnetic field, once set, for all commonly used radicals. The variable power (CW or pulsed) output from the EIK is transmitted to the DNP-NMR probe using a quasi-optic system with a high power isolator and a corrugated waveguide which feeds the microwaves into the DNP-NMR probe. Curved mirrors inside the probe project the microwaves down the axis of the MAS rotor, giving a very efficient system such that maximum DNP enhancement is achieved with less than 3 W output from the microwave source. The DNP-NMR probe operates with a sample temperature down to 90 K whilst spinning at 8 kHz. Significant enhancements, in excess of 100 for bacteriorhodopsin in purple membrane (bR in PM), are shown along with spectra which are enhanced by ≈25 with respect to room temperature, for both the pharmaceutical furosemide and surface functionalised silica. These enhancements allow hitherto prohibitively time consuming experiments to be undertaken. The power at which the DNP enhancement in bR in PM saturates does not change significantly between 90 K and 170 K even though the enhancement drops by a factor of ≈11. As the DNP build up time decreases by a factor 3 over this temperature range, the reduction in T1n is presumably a significant contribution to the drop in enhancement.

  9. Re-introduction of transmembrane serine residues reduce the minimum pore diameter of channelrhodopsin-2.

    Directory of Open Access Journals (Sweden)

    Ryan Richards

    Full Text Available Channelrhodopsin-2 (ChR2 is a microbial-type rhodopsin found in the green algae Chlamydomonas reinhardtii. Under physiological conditions, ChR2 is an inwardly rectifying cation channel that permeates a wide range of mono- and divalent cations. Although this protein shares a high sequence homology with other microbial-type rhodopsins, which are ion pumps, ChR2 is an ion channel. A sequence alignment of ChR2 with bacteriorhodopsin, a proton pump, reveals that ChR2 lacks specific motifs and residues, such as serine and threonine, known to contribute to non-covalent interactions within transmembrane domains. We hypothesized that reintroduction of the eight transmembrane serine residues present in bacteriorhodopsin, but not in ChR2, will restrict the conformational flexibility and reduce the pore diameter of ChR2. In this work, eight single serine mutations were created at homologous positions in ChR2. Additionally, an endogenous transmembrane serine was replaced with alanine. We measured kinetics, changes in reversal potential, and permeability ratios in different alkali metal solutions using two-electrode voltage clamp. Applying excluded volume theory, we calculated the minimum pore diameter of ChR2 constructs. An analysis of the results from our experiments show that reintroducing serine residues into the transmembrane domain of ChR2 can restrict the minimum pore diameter through inter- and intrahelical hydrogen bonds while the removal of a transmembrane serine results in a larger pore diameter. Therefore, multiple positions along the intracellular side of the transmembrane domains contribute to the cation permeability of ChR2.

  10. Resolution of ion translocating proteolipid subclasses active in bacterial calcification

    International Nuclear Information System (INIS)

    Formation of calcium hydroxyapatite occurs on membrane surfaces via interaction of calcium, inorganic phosphate, phospholipids, calcifiable proteolipids, and ion flux to and from the nucleating site. Recently, this laboratory reported that proteolipids from the calcifying bacterium, Bacterionema matruchotti, act as an ionophore when reconstituted into bacteriorhodopsin-proteoliposomes. This ionophoric activity is blocked by [14C]dicyclohexylcarbodiimide ([14C]DCCD). SDS-PAGE shows that [14C]DCCD binds to a single band of Mr 8500. To determine whether proteins other than the [14C]DCCD-binding protein are involved, we examined the function of proteolipid species extracted by solvents of differing polarity. Proteolipids were isolated independently from chloroform:methanol (2:1) and chloroform:methanol:HCl (200:100:1) extracts of the bacteria by Sephadex LH-20 chromatography and were electrophoresed on 12.5% acrylamide gels. The chloroform:methanol extract contained a major hand at Mr 10,000 that was not present in gels of proteolipid isolated by acidified solvent. Proteolipids extracted in chloroform:methanol:HCl included a broad band at Mr 8500, which co-migrated with the [14C] DCCD-binding protein. The rate and extent of proton translocation were not altered when either proteolipid extract was added individually to bacteriorhodopsin proteoliposomes. However, when proteolipids isolated from the chloroform:methanol and chloroform:methanol:HCl extracts were combined, the rate and extent of translocation were increased. These data demonstrate that at least two proteolipid proteins are necessary for ionophoric activity, the Mr 10,000 protein isolated by chloroform:methanol 2:1 and the [14C]DCCD-binding protein requiring acidified solvent for extraction

  11. Structural changes in the photoactive site of proteorhodopsin during the primary photoreaction.

    Science.gov (United States)

    Bergo, Vladislav; Amsden, Jason J; Spudich, Elena N; Spudich, John L; Rothschild, Kenneth J

    2004-07-20

    Proteorhodopsin (PR), found in marine gamma-proteobacteria, is a newly discovered light-driven proton pump similar to bacteriorhodopsin (BR). Because of the widespread distribution of proteobacteria in the worldwide oceanic waters, this pigment may contribute significantly to the global solar energy input in the biosphere. We examined structural changes that occur during the primary photoreaction (PR --> K) of wild-type pigment and two mutants using low-temperature FTIR difference spectroscopy. Several vibrations detected in the 3500-3700 cm(-1) region are assigned on the basis of H(2)O --> H(2)(18)O exchange to the perturbation of one or more internal water molecules. Substitution of the negatively charged Schiff base counterion, Asp97, with the neutral asparagine caused a downshift of the ethylenic (C=C) and Schiff base (C=N) stretching modes, in agreement with the 27 nm red shift of the visible lambda(max). However, this replacement did not alter the normal all-trans to 13-cis isomerization of the chromophore or the environment of the detected water molecule(s). In contrast, substitution of Asn230, which is in a position to interact with the Schiff base, with Ala induces a 5 nm red shift of the visible lambda(max) and alters the PR chromophore structure, its isomerization to K, and the environment of the detected internal water molecules. The combination of FTIR and site-directed mutagenesis establishes that both Asp97 and Asn230 are perturbed during the primary phototransition. The environment of Asn230 is further altered during the thermal decay of K. These results suggest that significant differences exist in the conformational changes which occur in the photoactive sites of proteorhodopsin and bacteriorhodopsin during the primary photoreaction. PMID:15248764

  12. Ionic Polymer Microactuator Activated by Photoresponsive Organic Proton Pumps

    Directory of Open Access Journals (Sweden)

    Khaled M. Al-Aribe

    2015-10-01

    Full Text Available An ionic polymer microactuator driven by an organic photoelectric proton pump transducer is described in this paper. The light responsive transducer is fabricated by using molecular self-assembly to immobilize oriented bacteriorhodopsin purple membrane (PM patches on a bio-functionalized porous anodic alumina (PAA substrate. When exposed to visible light, the PM proton pumps produce a unidirectional flow of ions through the structure’s nano-pores and alter the pH of the working solution in a microfluidic device. The change in pH is sufficient to generate an osmotic pressure difference across a hydroxyethyl methacrylate-acrylic acid (HEMA-AA actuator shell and induce volume expansion or contraction. Experiments show that the transducer can generate an ionic gradient of 2.5 μM and ionic potential of 25 mV, producing a pH increase of 0.42 in the working solution. The ΔpH is sufficient to increase the volume of the HEMA-AA microactuator by 80%. The volumetric transformation of the hydrogel can be used as a valve to close a fluid transport micro-channel or apply minute force to a mechanically flexible microcantilever beam.

  13. Time-resolved structural studies with serial crystallography: A new light on retinal proteins

    Directory of Open Access Journals (Sweden)

    Valérie Panneels

    2015-07-01

    Full Text Available Structural information of the different conformational states of the two prototypical light-sensitive membrane proteins, bacteriorhodopsin and rhodopsin, has been obtained in the past by X-ray cryo-crystallography and cryo-electron microscopy. However, these methods do not allow for the structure determination of most intermediate conformations. Recently, the potential of X-Ray Free Electron Lasers (X-FELs for tracking the dynamics of light-triggered processes by pump-probe serial femtosecond crystallography has been demonstrated using 3D-micron-sized crystals. In addition, X-FELs provide new opportunities for protein 2D-crystal diffraction, which would allow to observe the course of conformational changes of membrane proteins in a close-to-physiological lipid bilayer environment. Here, we describe the strategies towards structural dynamic studies of retinal proteins at room temperature, using injector or fixed-target based serial femtosecond crystallography at X-FELs. Thanks to recent progress especially in sample delivery methods, serial crystallography is now also feasible at synchrotron X-ray sources, thus expanding the possibilities for time-resolved structure determination.

  14. Nanomechanical properties of proteins and membranes depend on loading rate and electrostatic interactions.

    Science.gov (United States)

    Medalsy, Izhar D; Müller, Daniel J

    2013-03-26

    Knowing the dynamic mechanical response of tissue, cells, membranes, proteins, nucleic acids, and carbohydrates to external perturbations is important to understand various biological and biotechnological problems. Atomic force microscopy (AFM)-based approaches are the most frequently used nanotechnologies to determine the mechanical properties of biological samples that range in size from microscopic to (sub)nanoscopic. However, the dynamic nature of biomechanical properties has barely been addressed by AFM imaging. In this work, we characterizethe viscoelastic properties of the native light-driven proton pump bacteriorhodopsin of the purple membrane of Halobacterium salinarum. Using force-distance curve (F-D)-based AFM we imaged purple membranes while force probing their mechanical response over a wide range of loading rates (from ∼0.5 to 100 μN/s). Our results show that the mechanical stiffness of protein and membrane increases with the loading rate up to a factor of 10 (from ∼0.3 to 3.2 N/m). In addition, the electrostatic repulsion between AFM tip and sample can alter the mechanical stiffness measured by AFM up to ∼60% (from ∼0.8 to 1.3 N/m).These findings indicate that the mechanical response of membranes and proteins and probably of other biomolecular systems should be determined at different loading rates to fully understand their properties. PMID:23442147

  15. Ion-Pumping Microbial Rhodopsins

    Directory of Open Access Journals (Sweden)

    Hideki eKandori

    2015-09-01

    Full Text Available Rhodopsins are light-sensing proteins used in optogenetics. The word rhodopsin originates from the Greek words rhodo and opsis, indicating rose and sight, respectively. Although the classical meaning of rhodopsin is the red-colored pigment in our eyes, the modern meaning of rhodopsin encompasses photoactive proteins containing a retinal chromophore in animals and microbes. Animal and microbial rhodopsins possess 11-cis and all-trans retinal, respectively, to capture light in seven transmembrane α-helices, and photoisomerizations into all-trans and 13-cis forms, respectively, initiate each function. Ion-transporting proteins can be found in microbial rhodopsins, such as light-gated channels and light-driven pumps, which are the main tools in optogenetics. Light-driven pumps, such as archaeal H+ pump bacteriorhodopsin (BR and Cl- pump halorhodopsin (HR, were discovered in the 1970s, and their mechanism has been extensively studied. On the other hand, different kinds of H+ and Cl- pumps have been found in marine bacteria, such as proteorhodopsin (PR and Fulvimarina pelagi rhodopsin (FR, respectively. In addition, a light-driven Na+ pump was found, Krokinobacter eikastus rhodopsin 2 (KR2. These light-driven ion-pumping microbial rhodopsins are classified as DTD, TSA, DTE, NTQ and NDQ rhodopsins for BR, HR, PR, FR and KR2, respectively. Recent understanding of ion-pumping microbial rhodopsins is reviewed in this paper.

  16. Photoreactions and Structural Changes of Anabaena Sensory Rhodopsin

    Directory of Open Access Journals (Sweden)

    Akira Kawanabe

    2009-12-01

    Full Text Available Anabaena sensory rhodopsin (ASR is an archaeal-type rhodopsin found in eubacteria. The gene encoding ASR forms a single operon with ASRT (ASR transducer which is a 14 kDa soluble protein, suggesting that ASR functions as a photochromic sensor by activating the soluble transducer. This article reviews the detailed photoreaction processes of ASR, which were studied by low-temperature Fourier-transform infrared (FTIR and UV-visible spectroscopy. The former research reveals that the retinal isomerization is similar to bacteriorhodopsin (BR, but the hydrogen-bonding network around the Schiff base and cytoplasmic region is different. The latter study shows the stable photoproduct of the all-trans form is 100% 13-cis, and that of the 13-cis form is 100% all-trans. These results suggest that the structural changes of ASR in the cytoplasmic domain play important roles in the activation of the transducer protein, and photochromic reaction is optimized for its sensor function.

  17. Archaebacterial rhodopsin sequences: Implications for evolution

    Science.gov (United States)

    Lanyi, J. K.

    1991-01-01

    It was proposed over 10 years ago that the archaebacteria represent a separate kingdom which diverged very early from the eubacteria and eukaryotes. It follows that investigations of archaebacterial characteristics might reveal features of early evolution. So far, two genes, one for bacteriorhodopsin and another for halorhodopsin, both from Halobacterium halobium, have been sequenced. We cloned and sequenced the gene coding for the polypeptide of another one of these rhodopsins, a halorhodopsin in Natronobacterium pharaonis. Peptide sequencing of cyanogen bromide fragments, and immuno-reactions of the protein and synthetic peptides derived from the C-terminal gene sequence, confirmed that the open reading frame was the structural gene for the pharaonis halorhodopsin polypeptide. The flanking DNA sequences of this gene, as well as those of other bacterial rhodopsins, were compared to previously proposed archaebacterial consensus sequences. In pairwise comparisons of the open reading frame with DNA sequences for bacterio-opsin and halo-opsin from Halobacterium halobium, silent divergences were calculated. These indicate very considerable evolutionary distance between each pair of genes, even in the dame organism. In spite of this, three protein sequences show extensive similarities, indicating strong selective pressures.

  18. Fiscal 1998 research report on the production of bio-affinity materials by using microgravity environment; Bisho juryoku kankyo riyo seitai shinwa zairyo no sosei kenkyu 1998 nendo chosa hokokusho

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-03-01

    Experiment was made on production of bio-affinity materials under a microgravity condition by using the drop experiment facility of the underground a gravity experiment center. The experiment includes fast polymerization of polymer grains, protein orientation control and protein thin film formation, organic thin film formation, O/W emulsion preparation, and crystallization of protein under a microgravity condition. In polymerization of fine pyrrole grains under the existence of fine latex grains, production of fiber polypyrrole is remarkable, and fiber growth from latex surfaces was confirmed. The obtained thin bacteriorhodopsin film shows a photoelectric response, and as the AFM observation result, it is mainly composed of violet single-layer electrodeposited films. The wetting rise of acrylic monomer at the interface between solid and liquid is largely dependent on viscosity. The oil droplet size distribution of O/W emulsion is narrower than that in ground experiment, and is close to a normal distribution. Easy imaging of products was achieved by tracing cytochrome crystals with change in turbidity. The crystallization phase diagram of black mold acid protease A, and basic data in crystallization process were obtained. (NEDO)

  19. Modeling and design of light powered biomimicry micropump utilizing transporter proteins

    Science.gov (United States)

    Liu, Jin; Sze, Tsun-Kay Jackie; Dutta, Prashanta

    2014-11-01

    The creation of compact micropumps to provide steady flow has been an on-going challenge in the field of microfluidics. We present a mathematical model for a micropump utilizing Bacteriorhodopsin and sugar transporter proteins. This micropump utilizes transporter proteins as method to drive fluid flow by converting light energy into chemical potential. The fluid flow through a microchannel is simulated using the Nernst-Planck, Navier-Stokes, and continuity equations. Numerical results show that the micropump is capable of generating usable pressure. Designing parameters influencing the performance of the micropump are investigated including membrane fraction, lipid proton permeability, illumination, and channel height. The results show that there is a substantial membrane fraction region at which fluid flow is maximized. The use of lipids with low membrane proton permeability allows illumination to be used as a method to turn the pump on and off. This capability allows the micropump to be activated and shut off remotely without bulky support equipment. This modeling work provides new insights on mechanisms potentially useful for fluidic pumping in self-sustained bio-mimic microfluidic pumps. This work is supported in part by the National Science Fundation Grant CBET-1250107.

  20. Schlieren optics for leak detection

    Science.gov (United States)

    Peale, Robert E.; Ruffin, Alranzo B.

    1995-01-01

    The purpose of this research was to develop an optical method of leak detection. Various modifications of schlieren optics were explored with initial emphasis on leak detection of the plumbing within the orbital maneuvering system of the space shuttle (OMS pod). The schlieren scheme envisioned for OMS pod leak detection was that of a high contrast pattern on flexible reflecting material imaged onto a negative of the same pattern. We find that the OMS pod geometry constrains the characteristic length scale of the pattern to the order of 0.001 inch. Our experiments suggest that optical modulation transfer efficiency will be very low for such patterns, which will limit the sensitivity of the technique. Optical elements which allow a negative of the scene to be reversibly recorded using light from the scene itself were explored for their potential in adaptive single-ended schlieren systems. Elements studied include photochromic glass, bacteriorhodopsin, and a transmissive liquid crystal display. The dynamics of writing and reading patterns were studied using intensity profiles from recorded images. Schlieren detection of index gradients in air was demonstrated.

  1. Low-Z polymer sample supports for fixed-target serial femtosecond X-ray crystallography

    Energy Technology Data Exchange (ETDEWEB)

    Feld, Geoffrey K. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); National Institute of Environmental Health Science, Research Triangle Park, NC (United States); Heymann, Michael [Brandeis Univ., Waltham, MA (United States); Univ. of Hamburg and DESY, Hamburg (Germany); Benner, W. Henry [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Pardini, Tommaso [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Tsai, Ching -Ju [Paul Scherrer Inst. (PSI), Villigen (Switzerland); Boutet, Sebastien [SLAC National Accelerator Lab., Menlo Park, CA (United States); Coleman, Matthew A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Hunter, Mark S. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States); Li, Xiaodan [Paul Scherrer Inst. (PSI), Villigen (Switzerland); Messerschmidt, Marc [SLAC National Accelerator Lab., Menlo Park, CA (United States); BioXFEL Science and Technology Center, Buffalo, NY (United States); Opathalage, Achini [Brandeis Univ., Waltham, MA (United States); Pedrini, Bill [Paul Scherrer Inst. (PSI), Villigen (Switzerland); Williams, Garth J. [SLAC National Accelerator Lab., Menlo Park, CA (United States); Krantz, Bryan A. [Univ. of California, Berkeley, CA (United States); Fraden, Seth [Brandeis Univ., Waltham, MA (United States); Hau-Riege, Stefan [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Evans, James E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Segelke, Brent W. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Frank, Matthias [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-06-27

    X-ray free-electron lasers (XFELs) offer a new avenue to the structural probing of complex materials, including biomolecules. Delivery of precious sample to the XFEL beam is a key consideration, as the sample of interest must be serially replaced after each destructive pulse. The fixed-target approach to sample delivery involves depositing samples on a thin-film support and subsequent serial introduction via a translating stage. Some classes of biological materials, including two-dimensional protein crystals, must be introduced on fixed-target supports, as they require a flat surface to prevent sample wrinkling. A series of wafer and transmission electron microscopy (TEM)-style grid supports constructed of low-Z plastic have been custom-designed and produced. Aluminium TEM grid holders were engineered, capable of delivering up to 20 different conventional or plastic TEM grids using fixed-target stages available at the Linac Coherent Light Source (LCLS). As proof-of-principle, X-ray diffraction has been demonstrated from two-dimensional crystals of bacteriorhodopsin and three-dimensional crystals of anthrax toxin protective antigen mounted on these supports at the LCLS. In conclusion, the benefits and limitations of these low-Z fixed-target supports are discussed; it is the authors' belief that they represent a viable and efficient alternative to previously reported fixed-target supports for conducting diffraction studies with XFELs.

  2. Dynamic force microscopy imaging of native membranes

    International Nuclear Information System (INIS)

    We employed magnetic ACmode atomic force microscopy (MACmode AFM) as a novel dynamic force microscopy method to image surfaces of biological membranes in their native environments. The lateral resolution achieved under optimized imaging conditions was in the nanometer range, even when the sample was only weakly attached to the support. Purple membranes (PM) from Halobacterium salinarum were used as a test standard for topographical imaging. The hexagonal arrangement of the bacteriorhodopsin trimers on the cytoplasmic side of PM was resolved with 1.5 nm lateral accuracy, a resolution similar to images obtained in contact and tapping-mode AFM. Human rhinovirus 2 (HRV2) particles were attached to mica surfaces via nonspecific interactions. The capsid structure and 2 nm sized protein loops of HRV2 were routinely obtained without any displacement of the virus. Globular and filamentous structures on living and fixed endothelial cells were observed with a resolution of 5-20 nm. These examples show that MACmode AFM is a favorable method in studying the topography of soft and weakly attached biological samples with high resolution under physiological conditions

  3. The study of terbium regenerated bacterirhodopsin

    Institute of Scientific and Technical Information of China (English)

    XU; Bing; ZHANG; Yue; HU; Kunsheng

    2005-01-01

    The localization of Terbium (Tb3+) cations binding to deionized bacteriorhodopsin (bR) has been studied by using spectroscopic methods. It was found that adding Tb3+ cations to deionized bR affects the fluorescence lifetimes of tryptophan (Trp) in bR, the wavelength of fluorescence peak shifts "blue" and the peak value of fluorescence decreases. It was also found that adding one Tb3+ cation to deionized bR can restore the purple state from its blue state obviously. The measurements of absorbance, fluorescence and lifetime of fluorescence also show that when more than three Tb3+ cations are added, no further changes can be found. It is suggested that one Tb3+ specific binding site for the color-controlling is located on the exterior of the bR trimer structure to negatively charged lipids near Trp-10 and Trp-12. Three Tb3+ cations binding per bR is needed for the regenerated bR.

  4. Predictive energy landscapes for folding membrane protein assemblies.

    Science.gov (United States)

    Truong, Ha H; Kim, Bobby L; Schafer, Nicholas P; Wolynes, Peter G

    2015-12-28

    We study the energy landscapes for membrane protein oligomerization using the Associative memory, Water mediated, Structure and Energy Model with an implicit membrane potential (AWSEM-membrane), a coarse-grained molecular dynamics model previously optimized under the assumption that the energy landscapes for folding α-helical membrane protein monomers are funneled once their native topology within the membrane is established. In this study we show that the AWSEM-membrane force field is able to sample near native binding interfaces of several oligomeric systems. By predicting candidate structures using simulated annealing, we further show that degeneracies in predicting structures of membrane protein monomers are generally resolved in the folding of the higher order assemblies as is the case in the assemblies of both nicotinic acetylcholine receptor and V-type Na(+)-ATPase dimers. The physics of the phenomenon resembles domain swapping, which is consistent with the landscape following the principle of minimal frustration. We revisit also the classic Khorana study of the reconstitution of bacteriorhodopsin from its fragments, which is the close analogue of the early Anfinsen experiment on globular proteins. Here, we show the retinal cofactor likely plays a major role in selecting the final functional assembly. PMID:26723586

  5. Low power spatial light modulator with pharaonis phoborhodopsin

    International Nuclear Information System (INIS)

    Spatial light modulation (SLM) has been theoretically analysed in pharaonis phoborhodopsin (ppR) and its mutants based on nonlinear intensity induced excited-state absorption, to achieve large percentage modulation at low power. Amplitude modulation of probe laser read beam (I p') transmissions at 560, 512 and 390 nm, corresponding to the peak absorption of ppRO, ppRKL and ppRM intermediate states, respectively, of ppR photocycle, by write beam intensity (I m') at 498 nm, corresponding to the peak absorption of the initial ppR state, have been analysed using the rate equation approach, considering all six intermediate states in its photocycle. The SLM characteristics are shown to be sensitive to the normalized small signal absorption coefficient (β) and the rate constants of intermediates. For a given I m' range, there is an optimum value of β (β opt) for maximum percentage modulation. We can achieve 100% modulation of the read beam if the initial ppR state does not absorb the respective probe beams. The SLM characteristics of F86D ppR have also been used to design an all-optical XOR logic gate. High dynamic range and sensitivity can be achieved at low write beam intensities in ppR compared to wild-type bacteriorhodopsin (WT-bR)

  6. A network model to correlate conformational change and the impedance spectrum of single proteins

    Science.gov (United States)

    Alfinito, Eleonora; Pennetta, Cecilia; Reggiani, Lino

    2008-02-01

    Integrated nanodevices based on proteins or biomolecules are attracting increasing interest in today's research. In fact, it has been shown that proteins such as azurin and bacteriorhodopsin manifest some electrical properties that are promising for the development of active components of molecular electronic devices. Here we focus on two relevant kinds of protein: bovine rhodopsin, prototype of G-protein-coupled-receptor (GPCR) proteins, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer's disease. Both these proteins exert their function starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different impedance spectra associated with the different configurations. The distinct types of conformational change of rhodopsin and AChE agree with their dissimilar electrical responses. In particular, for rhodopsin the model predicts variations of the impedance spectra up to about 30%, while for AChE the same variations are limited to about 10%, which supports the existence of a dynamical equilibrium between its native and complexed states.

  7. A network model to correlate conformational change and the impedance spectrum of single proteins

    Energy Technology Data Exchange (ETDEWEB)

    Alfinito, Eleonora; Pennetta, Cecilia; Reggiani, Lino [Dipartimento di Ingegneria dell' Innovazione, Universita del Salento, Via Arnesano, Lecce (Italy); Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia (CNISM) (Italy)

    2008-02-13

    Integrated nanodevices based on proteins or biomolecules are attracting increasing interest in today's research. In fact, it has been shown that proteins such as azurin and bacteriorhodopsin manifest some electrical properties that are promising for the development of active components of molecular electronic devices. Here we focus on two relevant kinds of protein: bovine rhodopsin, prototype of G-protein-coupled-receptor (GPCR) proteins, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer's disease. Both these proteins exert their function starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different impedance spectra associated with the different configurations. The distinct types of conformational change of rhodopsin and AChE agree with their dissimilar electrical responses. In particular, for rhodopsin the model predicts variations of the impedance spectra up to about 30%, while for AChE the same variations are limited to about 10%, which supports the existence of a dynamical equilibrium between its native and complexed states.

  8. Photoactive mitochondria: in vivo transfer of a light-driven proton pump into the inner mitochondrial membrane of Schizosaccharomyces pombe.

    Science.gov (United States)

    Hoffmann, A; Hildebrandt, V; Heberle, J; Büldt, G

    1994-09-27

    The light-driven proton pump bacteriorhodopsin (bR) from Halobacterium salinarium has been genetically transferred into the inner mitochondrial membrane (IM) of the eukaryotic cell Schizosaccharomyces pombe, where the archaebacterial proton pump replaces or increases the proton gradient usually formed by the respiratory chain. For targeting and integration, as well as for the correct orientation of bR in the IM, the bacterioopsin gene (bop) was fused to signal sequences of IM proteins. Northern and Western blot analysis proved that all hybrid gene constructs containing the bop gene and a mitochondrial signal sequence were expressed and processed to mature bR. Fast transient absorption spectroscopy showed photocycle activity of bR integrated in the IM by formation of the M intermediate. Experiments with the pH-sensitive fluorescence dye 2',7'-bis(2-carboxyethyl)-5 (and -6)-carboxyfluorescein revealed bR-mediated proton pumping from the mitochondrial matrix into the intermembrane space. Glucose uptake measurements under anaerobic conditions showed that yeast cells containing photoactive mitochondria need less sugar under illumination. In summary, our experiments demonstrate the functional genetic transfer of a light energy converter to a naturally nonphotoactive eukaryotic organism. PMID:7937771

  9. High-throughput single-molecule force spectroscopy for membrane proteins

    Energy Technology Data Exchange (ETDEWEB)

    Bosshart, Patrick D; Casagrande, Fabio; Frederix, Patrick L T M; Engel, Andreas; Fotiadis, Dimitrios [M E Mueller Institute for Structural Biology, Biozentrum of the University of Basel, CH-4056 Basel (Switzerland); Ratera, Merce; Palacin, Manuel [Institute for Research in Biomedicine, Barcelona Science Park, Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona and Centro de Investigacion Biomedica en Red de Enfermedades Raras, E-08028 Barcelona (Spain); Bippes, Christian A; Mueller, Daniel J [BioTechnology Center, Technical University, Tatzberg 47, D-01307 Dresden (Germany)], E-mail: andreas.engel@unibas.ch, E-mail: dimitrios.fotiadis@mci.unibe.ch

    2008-09-24

    Atomic force microscopy-based single-molecule force spectroscopy (SMFS) is a powerful tool for studying the mechanical properties, intermolecular and intramolecular interactions, unfolding pathways, and energy landscapes of membrane proteins. One limiting factor for the large-scale applicability of SMFS on membrane proteins is its low efficiency in data acquisition. We have developed a semi-automated high-throughput SMFS (HT-SMFS) procedure for efficient data acquisition. In addition, we present a coarse filter to efficiently extract protein unfolding events from large data sets. The HT-SMFS procedure and the coarse filter were validated using the proton pump bacteriorhodopsin (BR) from Halobacterium salinarum and the L-arginine/agmatine antiporter AdiC from the bacterium Escherichia coli. To screen for molecular interactions between AdiC and its substrates, we recorded data sets in the absence and in the presence of L-arginine, D-arginine, and agmatine. Altogether {approx}400 000 force-distance curves were recorded. Application of coarse filtering to this wealth of data yielded six data sets with {approx}200 (AdiC) and {approx}400 (BR) force-distance spectra in each. Importantly, the raw data for most of these data sets were acquired in one to two days, opening new perspectives for HT-SMFS applications.

  10. Water-containing hydrogen-bonding network in the active center of channelrhodopsin.

    Science.gov (United States)

    Ito, Shota; Kato, Hideaki E; Taniguchi, Reiya; Iwata, Tatsuya; Nureki, Osamu; Kandori, Hideki

    2014-03-01

    Channelrhodopsin (ChR) functions as a light-gated ion channel in Chlamydomonas reinhardtii. Passive transport of cations by ChR is fundamentally different from the active transport by light-driven ion pumps such as archaerhodopsin, bacteriorhodopsin, and halorhodopsin. These microbial rhodopsins are important tools for optogenetics, where ChR is used to activate neurons by light, while the ion pumps are used for neural silencing. Ion-transport functions by these rhodopsins strongly depend on the specific hydrogen-bonding networks containing water near the retinal chromophore. In this work, we measured protein-bound water molecules in a chimeric ChR protein of ChR1 (helices A to E) and ChR2 (helices F and G) of Chlamydomonas reinhardtii using low-temperature FTIR spectroscopy at 77 K. We found that the active center of ChR possesses more water molecules (9 water vibrations) than those of other microbial (2-6 water vibrations) and animal (6-8 water vibrations) rhodopsins. We conclude that the protonated retinal Schiff base interacts with the counterion (Glu162) directly, without the intervening water molecule found in proton-pumping microbial rhodopsins. The present FTIR results and the recent X-ray structure of ChR reveal a unique hydrogen-bonding network around the active center of this light-gated ion channel. PMID:24512107

  11. Investigation of evolution-related aspects of bacterial rhodopsins

    Science.gov (United States)

    1994-01-01

    We have investigated evolution-related aspects of bacterial rhodopsins, the unique retinal-based energy transducing systems of halophilic archae. The approach was to describe both structural and functional aspects: the structure by sequencing genes to explore which regions are conserved, and the function by comparing proton and chloride transport in the closely related systems, bacteriorhodopsin and halorhodopsin, respectively. In the latter, we have made a good start toward the ultimate goal of separating the attributes of the general principles of retinal-based ionic pumps from those of the specific ion specificities, by determining the thermodynamics of the internal steps of the protein-mediated active transport process, as well as some of the intraprotein ion-transfer steps. Our present emphasis is on continuing to acquire the tools for studying what distinguishes proton transport from chloride transport. We consider it important, therefore, that we have been able to provide firm mathematical grounds for the kinetics analyses which underlies these studies. Our molecular biological studies have received a great boost from the expression vector for the bop gene based on a halobacterial plasmid, that we recently developed.

  12. Halophilic archaea on Earth and in space: growth and survival under extreme conditions.

    Science.gov (United States)

    Oren, Aharon

    2014-12-13

    Salts are abundant on Mars, and any liquid water that is present or may have been present on the planet is expected to be hypersaline. Halophilic archaea (family Halobacteriaceae) are the microorganisms best adapted to life at extremes of salinity on Earth. This paper reviews the properties of the Halobacteriaceae that may make the group good candidates for life also on Mars. Many species resist high UV and gamma radiation levels; one species has survived exposure to vacuum and radiation during a space flight; and there is at least one psychrotolerant species. Halophilic archaea may survive for millions of years within brine inclusions in salt crystals. Many species have different modes of anaerobic metabolism, and some can use light as an energy source using the light-driven proton pump bacteriorhodopsin. They are also highly tolerant to perchlorate, recently shown to be present in Martian soils, and some species can even use perchlorate as an electron acceptor to support anaerobic growth. The presence of characteristic carotenoid pigments (α-bacterioruberin and derivatives) makes the Halobacteriaceae easy to identify by Raman spectroscopy. Thus, if present on Mars, such organisms may be detected by Raman instrumentation planned to explore Mars during the upcoming ExoMars mission. PMID:25368347

  13. Super-Sensitive and Robust Biosensors from Supported Polymer Bilayers

    Energy Technology Data Exchange (ETDEWEB)

    Paxton, Walter F. [Sandia National Laboratories (SNL-NM), Albuquerque, NM (United States)

    2015-09-01

    Biological organisms are potentially the most sensitive and selective biological detection systems known, yet we are currently severely limited in our ability to exploit biological interactions in sensory devices, due in part to the limited stability of biological systems and derived materials. This proposal addresses an important aspect of integrating biological sensory materials in a solid state device. If successful, such technology could enable entirely new classes of robust biosensors that could be miniaturized and deployed in the field. The critical aims of the proposed work were 1) the calibration of a more versatile approach to measuring pH, 2) the use of this method to monitor pH changes caused by the light-induced pumping of protons across vesicles with bacteriorhodopsin integrated into the membranes (either polymer or lipid); 3) the preparation of bilayer assemblies on platinum surfaces; 4) the enhanced detection of lightinduced pH changes driven by bR-loaded supported bilayers. I have developed a methodology that may enable that at interfaces and developed a methodology to characterize the functionality of bilayer membranes with reconstituted membrane proteins. The integrity of the supported bilayer films however must be optimized prior to the full realization of the work originally envisioned in the original proposal. Nevertheless, the work performed on this project and the encouraging results it has produced has demonstrated that these goals are challenging yet within reach.

  14. Comparison of prokaryotic community structure from Mediterranean and Atlantic saltern concentrator ponds by a metagenomic approach

    Directory of Open Access Journals (Sweden)

    Ana Beatriz eFernández

    2014-05-01

    Full Text Available We analyzed the prokaryotic community structure of a saltern pond with 21 % total salts located in Isla Cristina, Huelva, Southwest Spain, close to the Atlantic ocean coast. For this purpose, we constructed a metagenome (designated as IC21 obtained by pyrosequencing consisting of 486 Mb with an average read length of 397 bp and compared it with other metagenomic datasets obtained from ponds with 19, 33 and 37 % total salts acquired from Santa Pola marine saltern, located in Alicante, East Spain, on the Mediterranean coast. Although the salinity in IC21 is closer to the pond with 19 % total salts from Santa Pola saltern (designated as SS19, IC21 is more similar at higher taxonomic levels to the pond with 33 % total salts from Santa Pola saltern (designated as SS33, since both are predominated by the phylum Euryarchaeota. However, there are significant differences at lower taxonomic levels where most sequences were related to the genus Halorubrum in IC21 and to Haloquadratum in SS33. Within the Bacteroidetes, the genus Psychroflexus is the most abundant in IC21 while Salinibacter dominates in SS33. Sequences related to bacteriorhodopsins and halorhodopsins correlate with the abundance of Haloquadratum in Santa Pola SS19 to SS33 and of Halorubrum in Isla Cristina IC21 dataset, respectively. Differences in composition might be attributed to local ecological conditions since IC21 showed a decrease in the number of sequences related to the synthesis of compatible solutes and in the utilization of phosphonate.

  15. Chirality in Nonlinear Optics

    Science.gov (United States)

    Haupert, Levi M.; Simpson, Garth J.

    2009-05-01

    The past decade has witnessed the emergence of new measurement approaches and applications for chiral thin films and materials enabled by the observations of the high sensitivity of second-order nonlinear optical measurements to chirality. In thin films, the chiral response to second harmonic generation and sum frequency generation (SFG) from a single molecular monolayer is often comparable with the achiral response. The chiral specificity also allows for symmetry-allowed SFG in isotropic chiral media, confirming predictions made ˜50 years ago. With these experimental demonstrations in hand, an important challenge is the construction of intuitive predictive models that allow the measured chiral response to be meaningfully related back to molecular and macromolecular structure. This review defines and considers three distinct mechanisms for chiral effects in uniaxially oriented assemblies: orientational chirality, intrinsic chirality, and isotropic chirality. The role of each is discussed in experimental and computational studies of bacteriorhodopsin films, binaphthol, and collagen. Collectively, these three model systems support a remarkably simple framework for quantitatively recovering the measured chiral-specific activity.

  16. Optical Pattern Recognition

    Science.gov (United States)

    Yu, Francis T. S.; Jutamulia, Suganda

    2008-10-01

    Contributors; Preface; 1. Pattern recognition with optics Francis T. S. Yu and Don A. Gregory; 2. Hybrid neural networks for nonlinear pattern recognition Taiwei Lu; 3. Wavelets, optics, and pattern recognition Yao Li and Yunglong Sheng; 4. Applications of the fractional Fourier transform to optical pattern recognition David Mendlovic, Zeev Zalesky and Haldum M. Oxaktas; 5. Optical implementation of mathematical morphology Tien-Hsin Chao; 6. Nonlinear optical correlators with improved discrimination capability for object location and recognition Leonid P. Yaroslavsky; 7. Distortion-invariant quadratic filters Gregory Gheen; 8. Composite filter synthesis as applied to pattern recognition Shizhou Yin and Guowen Lu; 9. Iterative procedures in electro-optical pattern recognition Joseph Shamir; 10. Optoelectronic hybrid system for three-dimensional object pattern recognition Guoguang Mu, Mingzhe Lu and Ying Sun; 11. Applications of photrefractive devices in optical pattern recognition Ziangyang Yang; 12. Optical pattern recognition with microlasers Eung-Gi Paek; 13. Optical properties and applications of bacteriorhodopsin Q. Wang Song and Yu-He Zhang; 14. Liquid-crystal spatial light modulators Aris Tanone and Suganda Jutamulia; 15. Representations of fully complex functions on real-time spatial light modulators Robert W. Cohn and Laurence G. Hassbrook; Index.

  17. Second Harmonic Generation as a Nondestructive Readout of Optical (Photo(electro)chromic and Magnetic) Memories

    Science.gov (United States)

    Aktsipetrov, O.; Fedyanin, A.; Melnikov, A.; Mishina, E.; Murzina, T.

    1998-01-01

    Optical second harmonic generation (SHG) is suggested as a nonlinear-optical nondestructive readout of (thin) film-based optical memories. Systematic studies of photoinduced, electroinduced, and magnetoinduced variations in the anisotropic SHG intensity in (i) thin solid photo(electro)chromic films of bacteriorhodopsin, (ii) photosensitive Langmuir-Blodgett films of 4-nitro-4'-N-octadecylazobenzene, and (iii) rare-earth iron garnet films show sufficient modification of their nonlinear-optical properties by these external factors. These photo-, electro- and magnetomodified nonlinear-optical properties of thin films being used for recording the information can be easily distinguished by the SHG readout. The IR fundamental radiation used for the excitation of the SHG is shown not to disturb the information recorded in photo(electro)chromic memories. For, what is to our knowledge, the first time, the surface-bulk crossterms in the SHG intensity originating from the interference of the crystalline and magnetization induced second harmonic fields are observed in magnetic iron garnet films.

  18. Femtosecond stimulated Raman spectroscopy of ultrafast biophysical reaction dynamics

    Science.gov (United States)

    McCamant, David William

    2004-12-01

    I have developed the technique of femtosecond stimulated Raman spectroscopy (FSRS), which enables the rapid acquisition of vibrational spectra with optical excitation to S2 (1Bu +) the molecule relaxes to S1 in 160 fs where it undergoes rapid two-step IVR with 200- and 450-fs time constants. In later work, the FSRS spectrum of S2 beta-carotene was observed, which consists of three intense and broad bands at ˜1100, 1300 and 1650 cm-1 that exhibit kinetics matching the decay of the S2 near-infrared absorption. These data show that there is no additional intermediate 1B u- electronic state involved in the relaxation pathway of beta-carotene. FSRS was also used to study the photoisomerization dynamics in bacteriorhodopsin (bR). Spectra obtained during bR's excited state lifetime exhibit dispersive lineshapes at the ground-state frequencies that decay in 250 fs and are attributed to a nonlinear emission process. This relaxation is significantly faster than the decay of the stimulated emission (˜500 fs), indicating that the excited population moves away from the ground-state geometry in 250 fs. Spectral changes between 1.5 to 100 ps reveal that a significant fraction of the isomerization occurs on the ground state photoproduct surface. The many benefits FSRS will make it a valuable tool for vibrational spectroscopy of reaction dynamics in ultrafast photochemical and photophysical processes.

  19. Improvement of the diffraction efficiency and kinetics of holographic gratings in photochromic media by auxiliary light

    Science.gov (United States)

    Hampp, Norbert; Juchem, Thorsten

    2004-12-01

    Holographic gratings recorded in photochromic media often do not obtain the maximally achievable diffraction efficiency because of diminishing the fringe contrast caused, e.g., by a photochemically active readout beam or unequal intensities of object and reference waves. For nonreversible materials this problem causes a decrease in diffraction efficiency that is proportional to the signal-to-noise ratio (SNR). However, in nonlinear materials such as photochromic media, for which saturation effects need to be considered, an out-of-proportion decrease in the SNR results. It is shown that an overshooting peak during hologram growth, which then decays to a lower permanent level of diffraction, is an indicator for such a situation. Even a weak readout beam may cause such effects, which significantly affect the hologram kinetics. The observed overshooting diffraction efficiency may even be misinterpreted to be dependent on material properties. Experimental and theoretical proof that with low levels of auxiliary light this type of problem can be eliminated completely is presented. Throughout this research bacteriorhodopsin films were used, but the results are valid for photochromic media in general.

  20. Novel implementation of nonlinear joint transform correlators in optical security and validation

    Science.gov (United States)

    Weber, David C.; Trolinger, James D.

    1999-01-01

    Emblems using holograms or other diffractive devices have long been used to mark cards and other objects as a means of authentication. The effectiveness of such emblems as security devices is ultimately determined by the inspection system. Due to the expense and highly variable performance of the human inspector, automated machine reading devices are an attractive alternative for performing the verification task. An additional advantage of the machine reactor is that information regarding the card or its holder can be stored covertly. A security verification system is presented consisting of a holographic security emblem in which information is covertly stored and an automated reader based on a joint transform correlator. A holographic encoding method is used to produce an emblem that stores the required phase and/or amplitude information in the form of a complex, 3D diffraction pattern that can be interpreted only through the use of a second `key' hologram. The reader incorporates the use of the nonlinear material, bacteriorhodopsin, as a means of miniaturizing the system, reducing system cost, and improving system performance. Experimental results are presented that demonstrate the feasibility of the approach for security applications.

  1. Low power spatial light modulator with pharaonis phoborhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, Parag [Department of Physics and Computer Science, Dayalbagh Educational Institute (Deemed University), Agra 282005 (India); Roy, Sukhdev [Department of Physics and Computer Science, Dayalbagh Educational Institute (Deemed University), Agra 282005 (India)]. E-mail: sukhdevr@hotmail.com; Singh, C.P. [Department of Physics and Computer Science, Dayalbagh Educational Institute (Deemed University), Agra 282005 (India)

    2005-04-22

    Spatial light modulation (SLM) has been theoretically analysed in pharaonis phoborhodopsin (ppR) and its mutants based on nonlinear intensity induced excited-state absorption, to achieve large percentage modulation at low power. Amplitude modulation of probe laser read beam (I {sub p}') transmissions at 560, 512 and 390 nm, corresponding to the peak absorption of ppR{sub O}, ppR{sub KL} and ppR{sub M} intermediate states, respectively, of ppR photocycle, by write beam intensity (I {sub m}') at 498 nm, corresponding to the peak absorption of the initial ppR state, have been analysed using the rate equation approach, considering all six intermediate states in its photocycle. The SLM characteristics are shown to be sensitive to the normalized small signal absorption coefficient ({beta}) and the rate constants of intermediates. For a given I {sub m}' range, there is an optimum value of {beta} ({beta} {sub opt}) for maximum percentage modulation. We can achieve 100% modulation of the read beam if the initial ppR state does not absorb the respective probe beams. The SLM characteristics of F86D ppR have also been used to design an all-optical XOR logic gate. High dynamic range and sensitivity can be achieved at low write beam intensities in ppR compared to wild-type bacteriorhodopsin (WT-bR)

  2. Recording of dynamic gratings in the nonlinear optical coating of a planar waveguide

    Science.gov (United States)

    Kozhevnikov, N. M.; Korolev, A. E.; Koklyushkin, A. V.; Lipovskaya, M. Yu.; Nazarov, V. N.

    2003-04-01

    The possibility of controlled energy exchange between interfering waveguide modes in a singlemode planar waveguide with a nonlinear optical coating is analyzed. As the coating, a suspension of bacteriorhodopsin D96N was used, which makes it possible to realize two spectrally separated mechanisms of recording and controlling dynamic gratings, i.e., the spatial modulation of the trans-cis excitation rate and the spatial modulation of the cis-trans relaxation rate. The method of phase-modulated beams was used to implement the energy exchange. The dynamic gratings in the coating were recorded by using both radiation with a wavelength within the absorption band of the trans state (630 nm) and radiation with a wavelength within the absorption band of the cis state (440 nm). Efficient control of the energy exchange between the waveguide modes by means of uniform exposure of their interference region to radiation with another wavelength was observed. A completely integral geometrical layout for optically controlled energy exchange was realized. The results obtained are compared with known data on energy exchange between beams in the bulk of a similar nonlinear medium.

  3. Taking electrons out of bioelectronics: bioprotonic memories and enzymatic logic gates (Presentation Recording)

    Science.gov (United States)

    Rolandi, Marco

    2015-10-01

    In living systems, protonic and ionic currents are the basis for all information processing. As such, artificial devices based on protonic and ionic currents offer an exciting opportunity for bioelectronics. Proton transport in nature is important for ATP oxidative phosphorylation, the HCVN1 voltage gated proton channel, light activated proton pumping in bacteriorhodopsin, and the proton conducting single water file of the antibiotic gramicidin. In these systems, protons move along hydrogen bond networks formed by water and the hydrated biomolecules (proton wires). We have previously demonstrated complementary H+- and OH-- FETs with acid and base doped biopolymer proton wires and PdHx proton conducting contacts. Here, I will discuss proton-conducting devices based oh highly conductive proton wires that emulate brain synapses, display memristive behaviour, and are connected to form shift registries. Furthermore, I will present the integration of these devices with enzymatic logic gates for integrated biotic-abiotic protonic information processing. Preliminary results on using these devices to affect biological function will be discussed.

  4. Organization, Structure and Activity of Proteins in Monolayers

    Energy Technology Data Exchange (ETDEWEB)

    Boucher,J.; Trudel, E.; Methot, M.; Desmeules, P.; Salesse, C.

    2007-01-01

    Many different processes take place at the cell membrane interface. Indeed, for instance, ligands bind membrane proteins which in turn activate peripheral membrane proteins, some of which are enzymes whose action is also located at the membrane interface. Native cell membranes are difficult to use to gain information on the activity of individual proteins at the membrane interface because of the large number of different proteins involved in membranous processes. Model membrane systems, such as monolayers at the air-water interface, have thus been extensively used during the last 50 years to reconstitute proteins and to gain information on their organization, structure and activity in membranes. In the present paper, we review the recent work we have performed with membrane and peripheral proteins as well as enzymes in monolayers at the air-water interface. We show that the structure and orientation of gramicidin has been determined by combining different methods. Furthermore, we demonstrate that the secondary structure of rhodopsin and bacteriorhodopsin is indistinguishable from that in native membranes when appropriate conditions are used. We also show that the kinetics and extent of monolayer binding of myristoylated recoverin is much faster than that of the nonmyristoylated form and that this binding is highly favored by the presence polyunsaturated phospholipids. Moreover, we show that the use of fragments of RPE65 allow determine which region of this protein is most likely involved in membrane binding. Monomolecular films were also used to further understand the hydrolysis of organized phospholipids by phospholipases A2 and C.

  5. Crystal Structure of Phosphatidylglycerophosphatase (PGPase), a Putative Membrane-Bound Lipid Phosphatase, Reveals a Novel Binuclear Metal Binding Site and Two Proton Wires

    Energy Technology Data Exchange (ETDEWEB)

    Kumaran,D.; Bonnano, J.; Burley, S.; Swaminathan, S.

    2006-01-01

    Phosphatidylglycerophosphatase (PGPase), an enzyme involved in lipid metabolism, catalyzes formation of phosphatidylglycerol from phosphatidylglycerophosphate. Phosphatidylglycerol is a multifunctional phospholipid, found in the biological membranes of many organisms. Here, we report the crystal structure of Listeria monocytogenes PGPase at 1.8 Angstroms resolution. PGPase, an all-helical molecule, forms a homotetramer. Each protomer contains an independent active site with two metal ions, Ca{sup 2+} and Mg{sup 2+}, forming a hetero-binuclear center located in a hydrophilic cavity near the surface of the molecule. The binuclear center, conserved ligands, metal-bound water molecules, and an Asp-His dyad form the active site. The catalytic mechanism of this enzyme is likely to proceed via binuclear metal activated nucleophilic water. The binuclear metal-binding active-site environment of this structure should provide insights into substrate binding and metal-dependent catalysis. A long channel with inter-linked linear water chains, termed 'proton wires', is observed at the tetramer interface. Comparison of similar water chain structures in photosynthetic reaction centers (RCs), Cytochrome f, gramicidin, and bacteriorhodopsin, suggests that PGPase may conduct protons via proton wires.

  6. Extension of the FACTS Implicit Solvation Model to Membranes.

    Science.gov (United States)

    Carballo-Pacheco, Martín; Vancea, Ioan; Strodel, Birgit

    2014-08-12

    The generalized Born (GB) formalism can be used to model water as a dielectric continuum. Among the different implicit solvent models using the GB formalism, FACTS is one of the fastest. Here, we extend FACTS so that it can represent a membrane environment. This extension is accomplished by considering a position dependent dielectric constant and empirical surface tension parameter. For the calculation of the effective Born radii in different dielectric environments we present a parameter-free approximation to Kirkwood's equation, which uses the Born radii obtained with FACTS for the water environment as input. This approximation is tested for the calculation of self-free energies, pairwise interaction energies in solution and solvation free energies of complete protein conformations. The results compare well to those from the finite difference Poisson method. The new implicit membrane model is applied to estimate free energy insertion profiles of amino acid analogues and in molecular dynamics simulations of melittin, WALP23 and KALP23, glycophorin A, bacteriorhodopsin, and a Clc channel dimer. In all cases, the results agree qualitatively with experiments and explicit solvent simulations. Moreover, the implicit membrane model is only six times slower than a vacuum simulation. PMID:26588287

  7. Resonance energy transfer in nano-bio hybrid structures can be modulated by UV laser irradiation

    International Nuclear Information System (INIS)

    A method for targeted variation of the radiation properties of quantum dots (QDs) to control the efficiency of resonance energy transfer in nanocrystal assemblies and nano-bio hybrid materials has been developed. The method is based on strong ultraviolet (UV) laser irradiation of QDs and allows the extinction and luminescence spectra to be controlled and the luminescence quantum yield and decay kinetics to be varied. Water-soluble QDs have been synthesized and used for analyzing the effect of energy transfer from semiconductor nanocrystals on the photocycle of the photosensitive protein bacteriorhodopsin (bR) in bR–QD complexes. The UV irradiation mode has been selected in a way permitting the modulation of QD optical parameters without modification of their structure or physico-chemical properties. It is concluded that the QD interaction with bR accelerates its photocycle, but this acceleration is determined by electrostatic interactions, rather than Förster resonance energy transfer from QDs to bR. The method of UV laser irradiation of fluorescent semiconductor QDs has proven to be an efficient technique for variation of nanocrystal optical properties without affecting their structure, as well as for fine modulation of the energy transfer processes in the nanocrystal assemblies and nano-bio hybrid materials. (letter)

  8. 7 Å Resolution in Protein 2-Dimentional-Crystal X-Ray Diffraction at Linac Coherent Light Source

    Energy Technology Data Exchange (ETDEWEB)

    Pedrini, Bill; Tsai, Ching-Ju; Capitani, Guido; Padeste, Celestino; Hunter, Mark; Zatsepin, Nadia A.; Barty, Anton; Benner, Henry; Boutet, Sebastien; Feld, Geoffrey K.; Hau-Riege, Stefan; Kirian, Rick; Kupitz, Christopher; Messerschmidt, Marc; Ogren, John I.; Pardini, Tommaso; Segelke, Brent; Williams, Garth J.; Spence , John C.; Abela, Rafael; Coleman, Matthew A.; Evans, James E.; Schertler, Gebhard; Frank, Matthias; Li, Xiao-Dan

    2014-06-09

    Membrane proteins arranged as two-dimensional (2D) crystals in the lipid en- vironment provide close-to-physiological structural information, which is essential for understanding the molecular mechanisms of protein function. X-ray diffraction from individual 2D crystals did not represent a suitable investigation tool because of radiation damage. The recent availability of ultrashort pulses from X-ray Free Electron Lasers (X-FELs) has now provided a mean to outrun the damage. Here we report on measurements performed at the LCLS X-FEL on bacteriorhodopsin 2D crystals mounted on a solid support and kept at room temperature. By merg- ing data from about a dozen of single crystal diffraction images, we unambiguously identified the diffraction peaks to a resolution of 7 °A, thus improving the observable resolution with respect to that achievable from a single pattern alone. This indicates that a larger dataset will allow for reliable quantification of peak intensities, and in turn a corresponding increase of resolution. The presented results pave the way to further X-FEL studies on 2D crystals, which may include pump-probe experiments at subpicosecond time resolution.

  9. Ultrashort Phenomena in Biochemistry and Biological Signaling

    International Nuclear Information System (INIS)

    In biological phenomena there are indications that within the long pulse-length of the action potential on millisecond scale, there is additional ultrashort perturbation encoding that provides the brain with detailed information about the origin (location) and physiological characteristics. The objective is to identify the mechanism-of-action providing the potential for encoding in biological signal propagation. The actual molecular processes involved in the initiation of the action potential have been identified to be in the femtosecond and pico-second scale. The depolarization process of the cellular membrane itself, leading to the onset of the actionpotential that is transmitted to the brain, however is in the millisecond timeframe. One example of the femtosecond chemical interaction is the photoresponse of bacteriorhodopsin. No clear indication for the spatial encoding has so far been verified. Further research will be required on a cellular signal analysis level to confirm or deny the spatial and physiological encoding in the signal wave-trains of intercellular communications and sensory stimuli. The pathological encoding process for cardiac depolarization is however very pronounced and validated, however this electro-chemical process is in the millisecond amplitude and frequency modulation spectrum

  10. Protein cleavage strategies for an improved analysis of the membrane proteome

    Directory of Open Access Journals (Sweden)

    Poetsch Ansgar

    2006-03-01

    Full Text Available Abstract Background Membrane proteins still remain elusive in proteomic studies. This is in part due to the distribution of the amino acids lysine and arginine, which are less frequent in integral membrane proteins and almost absent in transmembrane helices. As these amino acids are cleavage targets for the commonly used protease trypsin, alternative cleavage conditions, which should improve membrane protein analysis, were tested by in silico digestion for the three organisms Saccharomyces cerevisiae, Halobacterium sp. NRC-1, and Corynebacterium glutamicum as hallmarks for eukaryotes, archea and eubacteria. Results For the membrane proteomes from all three analyzed organisms, we identified cleavage conditions that achieve better sequence and proteome coverage than trypsin. Greater improvement was obtained for bacteria than for yeast, which was attributed to differences in protein size and GRAVY. It was demonstrated for bacteriorhodopsin that the in silico predictions agree well with the experimental observations. Conclusion For all three examined organisms, it was found that a combination of chymotrypsin and staphylococcal peptidase I gave significantly better results than trypsin. As some of the improved cleavage conditions are not more elaborate than trypsin digestion and have been proven useful in practice, we suppose that the cleavage at both hydrophilic and hydrophobic amino acids should facilitate in general the analysis of membrane proteins for all organisms.

  11. Ultrashort Phenomena in Biochemistry and Biological Signaling

    Science.gov (United States)

    Splinter, Robert

    2014-11-01

    In biological phenomena there are indications that within the long pulse-length of the action potential on millisecond scale, there is additional ultrashort perturbation encoding that provides the brain with detailed information about the origin (location) and physiological characteristics. The objective is to identify the mechanism-of-action providing the potential for encoding in biological signal propagation. The actual molecular processes involved in the initiation of the action potential have been identified to be in the femtosecond and pico-second scale. The depolarization process of the cellular membrane itself, leading to the onset of the actionpotential that is transmitted to the brain, however is in the millisecond timeframe. One example of the femtosecond chemical interaction is the photoresponse of bacteriorhodopsin. No clear indication for the spatial encoding has so far been verified. Further research will be required on a cellular signal analysis level to confirm or deny the spatial and physiological encoding in the signal wave-trains of intercellular communications and sensory stimuli. The pathological encoding process for cardiac depolarization is however very pronounced and validated, however this electro-chemical process is in the millisecond amplitude and frequency modulation spectrum.

  12. Probing Saltern Brines with an Oxygen Electrode: What Can We Learn about the Community Metabolism in Hypersaline Systems?

    Science.gov (United States)

    Oren, Aharon

    2016-01-01

    We have explored the use of optical oxygen electrodes to study oxygenic photosynthesis and heterotrophic activities in crystallizer brines of the salterns in Eilat, Israel. Monitoring oxygen uptake rates in the dark enables the identification of organic substrates that are preferentially used by the community. Addition of glycerol (the osmotic solute synthesized by Dunaliella) or dihydroxyacetone (produced from glycerol by Salinibacter) enhanced respiration rates. Pyruvate, produced from glycerol or from some sugars by certain halophilic Archaea also stimulated community respiration. Fumarate had a sparing effect on respiration, possibly as many halophilic Archaea can use fumarate as a terminal electron acceptor in respiration. Calculating the photosynthetic activity of Dunaliella by monitoring oxygen concentration changes during light/dark incubations is not straightforward as light also affects respiration of some halophilic Archaea and Bacteria due to action of light-driven proton pumps. When illuminated, community respiration of brine samples in which oxygenic photosynthesis was inhibited by DCMU decreased by ~40%. This effect was interpreted as the result of competition between two energy yielding systems: the bacteriorhodopsin proton pump and the respiratory chain of the prokaryotes. These findings have important implications for the interpretation of other published data on photosynthetic and respiratory activities in hypersaline environments. PMID:27338478

  13. New ultrarapid-scanning interferometer for FT-IR spectroscopy with microsecond time-resolution

    Science.gov (United States)

    Süss, B.; Ringleb, F.; Heberle, J.

    2016-06-01

    A novel Fourier-transform infrared (FT-IR) rapid-scan spectrometer has been developed (patent pending EP14194520.4) which yields 1000 times higher time resolution as compared to conventional rapid-scanning spectrometers. The central element to achieve faster scanning rates is based on a sonotrode whose front face represents the movable mirror of the interferometer. A prototype spectrometer with a time resolution of 13 μs was realized, capable of fully automated long-term measurements with a flow cell for liquid samples, here a photosynthetic membrane protein in solution. The performance of this novel spectrometer is demonstrated by recording the photoreaction of bacteriorhodopsin initiated by a short laser pulse that is synchronized to the data recording. The resulting data are critically compared to those obtained by step-scan spectroscopy and demonstrate the relevance of performing experiments on proteins in solution. The spectrometer allows for future investigations of fast, non-repetitive processes, whose investigation is challenging to step-scan FT-IR spectroscopy.

  14. Driving electrocatalytic activity by interface electronic structure control in a metalloprotein hybrid catalyst for efficient hydrogen evolution.

    Science.gov (United States)

    Behera, Sushant Kumar; Deb, Pritam; Ghosh, Arghya

    2016-08-17

    The rational design of metalloprotein hybrid structures and precise calculations for understanding the role of the interfacial electronic structure in regulating the HER activity of water splitting sites and their microscopic effect for obtaining robust hydrogen evolution possess great promise for developing highly efficient nano-bio hybrid HER catalysts. Here, we employ high-accuracy linear-scaling density functional theory calculations using a near-complete basis set and a minimal parameter implicit solvent model within the self-consistent calculations, on silver (Ag) ions assimilated on bacteriorhodopsin (bR) at specific binding sites. Geometry optimization indicates the formation of active sites at the interface of the metalloprotein complex and the density of states reflects the metallic nature of the active sites. The reduced value of the canonical orbital gap indicates the state of dynamic nature after Ag ion assimilation on active sites and smooth electron transfer. These incorporated active protein sites are more efficient in electrolytic splitting of water than pristine sites due to their low value of Gibbs free energy for the HER in terms of hydrogen coverages. Volcano plot analysis and the free energy diagram are compared for understanding the hydrogen evolution efficiency. Moreover, the essential role of the interfacial electronic properties in regulating the HER catalytic activity of water splitting sites and enhancing the efficiency is elucidated. PMID:27499158

  15. Efficient unfolding pattern recognition in single molecule force spectroscopy data

    Directory of Open Access Journals (Sweden)

    Labudde Dirk

    2011-06-01

    Full Text Available Abstract Background Single-molecule force spectroscopy (SMFS is a technique that measures the force necessary to unfold a protein. SMFS experiments generate Force-Distance (F-D curves. A statistical analysis of a set of F-D curves reveals different unfolding pathways. Information on protein structure, conformation, functional states, and inter- and intra-molecular interactions can be derived. Results In the present work, we propose a pattern recognition algorithm and apply our algorithm to datasets from SMFS experiments on the membrane protein bacterioRhodopsin (bR. We discuss the unfolding pathways found in bR, which are characterised by main peaks and side peaks. A main peak is the result of the pairwise unfolding of the transmembrane helices. In contrast, a side peak is an unfolding event in the alpha-helix or other secondary structural element. The algorithm is capable of detecting side peaks along with main peaks. Therefore, we can detect the individual unfolding pathway as the sequence of events labeled with their occurrences and co-occurrences special to bR's unfolding pathway. We find that side peaks do not co-occur with one another in curves as frequently as main peaks do, which may imply a synergistic effect occurring between helices. While main peaks co-occur as pairs in at least 50% of curves, the side peaks co-occur with one another in less than 10% of curves. Moreover, the algorithm runtime scales well as the dataset size increases. Conclusions Our algorithm satisfies the requirements of an automated methodology that combines high accuracy with efficiency in analyzing SMFS datasets. The algorithm tackles the force spectroscopy analysis bottleneck leading to more consistent and reproducible results.

  16. Chloroflexi CL500-11 Populations That Predominate Deep-Lake Hypolimnion Bacterioplankton Rely on Nitrogen-Rich Dissolved Organic Matter Metabolism and C1 Compound Oxidation.

    Science.gov (United States)

    Denef, Vincent J; Mueller, Ryan S; Chiang, Edna; Liebig, James R; Vanderploeg, Henry A

    2016-03-01

    The Chloroflexi CL500-11 clade contributes a large proportion of the bacterial biomass in the oxygenated hypolimnia of deep lakes worldwide, including the world's largest freshwater system, the Laurentian Great Lakes. Traits that allow CL500-11 to thrive and its biogeochemical role in these environments are currently unknown. Here, we found that a CL500-11 population was present mostly in offshore waters along a transect in ultraoligotrophic Lake Michigan (a Laurentian Great Lake). It occurred throughout the water column in spring and only in the hypolimnion during summer stratification, contributing up to 18.1% of all cells. Genome reconstruction from metagenomic data suggested an aerobic, motile, heterotrophic lifestyle, with additional energy being gained through carboxidovory and methylovory. Comparisons to other available streamlined freshwater genomes revealed that the CL500-11 genome contained a disproportionate number of cell wall/capsule biosynthesis genes and the most diverse spectrum of genes involved in the uptake of dissolved organic matter (DOM) substrates, particularly peptides. In situ expression patterns indicated the importance of DOM uptake and protein/peptide turnover, as well as type I and type II carbon monoxide dehydrogenase and flagellar motility. Its location in the water column influenced its gene expression patterns the most. We observed increased bacteriorhodopsin gene expression and a response to oxidative stress in surface waters compared to its response in deep waters. While CL500-11 carries multiple adaptations to an oligotrophic lifestyle, its investment in motility, its large cell size, and its distribution in both oligotrophic and mesotrophic lakes indicate its ability to thrive under conditions where resources are more plentiful. Our data indicate that CL500-11 plays an important role in nitrogen-rich DOM mineralization in the extensive deep-lake hypolimnion habitat. PMID:26682860

  17. Integrated biclustering of heterogeneous genome-wide datasets for the inference of global regulatory networks

    Directory of Open Access Journals (Sweden)

    Baliga Nitin S

    2006-06-01

    Full Text Available Abstract Background The learning of global genetic regulatory networks from expression data is a severely under-constrained problem that is aided by reducing the dimensionality of the search space by means of clustering genes into putatively co-regulated groups, as opposed to those that are simply co-expressed. Be cause genes may be co-regulated only across a subset of all observed experimental conditions, biclustering (clustering of genes and conditions is more appropriate than standard clustering. Co-regulated genes are also often functionally (physically, spatially, genetically, and/or evolutionarily associated, and such a priori known or pre-computed associations can provide support for appropriately grouping genes. One important association is the presence of one or more common cis-regulatory motifs. In organisms where these motifs are not known, their de novo detection, integrated into the clustering algorithm, can help to guide the process towards more biologically parsimonious solutions. Results We have developed an algorithm, cMonkey, that detects putative co-regulated gene groupings by integrating the biclustering of gene expression data and various functional associations with the de novo detection of sequence motifs. Conclusion We have applied this procedure to the archaeon Halobacterium NRC-1, as part of our efforts to decipher its regulatory network. In addition, we used cMonkey on public data for three organisms in the other two domains of life: Helicobacter pylori, Saccharomyces cerevisiae, and Escherichia coli. The biclusters detected by cMonkey both recapitulated known biology and enabled novel predictions (some for Halobacterium were subsequently confirmed in the laboratory. For example, it identified the bacteriorhodopsin regulon, assigned additional genes to this regulon with apparently unrelated function, and detected its known promoter motif. We have performed a thorough comparison of cMonkey results against other

  18. Optimisation of NMR dynamic models II. A new methodology for the dual optimisation of the model-free parameters and the Brownian rotational diffusion tensor

    International Nuclear Information System (INIS)

    Finding the dynamics of an entire macromolecule is a complex problem as the model-free parameter values are intricately linked to the Brownian rotational diffusion of the molecule, mathematically through the autocorrelation function of the motion and statistically through model selection. The solution to this problem was formulated using set theory as an element of the universal set U-the union of all model-free spaces (d'Auvergne EJ and Gooley PR (2007) Mol BioSyst 3(7), 483-494). The current procedure commonly used to find the universal solution is to initially estimate the diffusion tensor parameters, to optimise the model-free parameters of numerous models, and then to choose the best model via model selection. The global model is then optimised and the procedure repeated until convergence. In this paper a new methodology is presented which takes a different approach to this diffusion seeded model-free paradigm. Rather than starting with the diffusion tensor this iterative protocol begins by optimising the model-free parameters in the absence of any global model parameters, selecting between all the model-free models, and finally optimising the diffusion tensor. The new model-free optimisation protocol will be validated using synthetic data from Schurr JM et al. (1994) J Magn Reson B 105(3), 211-224 and the relaxation data of the bacteriorhodopsin (1-36)BR fragment from Orekhov VY (1999) J Biomol NMR 14(4), 345-356. To demonstrate the importance of this new procedure the NMR relaxation data of the Olfactory Marker Protein (OMP) of Gitti R et al. (2005) Biochem 44(28), 9673-9679 is reanalysed. The result is that the dynamics for certain secondary structural elements is very different from those originally reported

  19. Proton transfers in a channelrhodopsin-1 studied by Fourier transform infrared (FTIR) difference spectroscopy and site-directed mutagenesis.

    Science.gov (United States)

    Ogren, John I; Yi, Adrian; Mamaev, Sergey; Li, Hai; Spudich, John L; Rothschild, Kenneth J

    2015-05-15

    Channelrhodopsin-1 from the alga Chlamydomonas augustae (CaChR1) is a low-efficiency light-activated cation channel that exhibits properties useful for optogenetic applications such as a slow light inactivation and a red-shifted visible absorption maximum as compared with the more extensively studied channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2). Previously, both resonance Raman and low-temperature FTIR difference spectroscopy revealed that unlike CrChR2, CaChR1 under our conditions exhibits an almost pure all-trans retinal composition in the unphotolyzed ground state and undergoes an all-trans to 13-cis isomerization during the primary phototransition typical of other microbial rhodopsins such as bacteriorhodopsin (BR). Here, we apply static and rapid-scan FTIR difference spectroscopy along with site-directed mutagenesis to characterize the proton transfer events occurring upon the formation of the long-lived conducting P2 (380) state of CaChR1. Assignment of carboxylic C=O stretch bands indicates that Asp-299 (homolog to Asp-212 in BR) becomes protonated and Asp-169 (homolog to Asp-85 in BR) undergoes a net change in hydrogen bonding relative to the unphotolyzed ground state of CaChR1. These data along with earlier FTIR measurements on the CaChR1 → P1 transition are consistent with a two-step proton relay mechanism that transfers a proton from Glu-169 to Asp-299 during the primary phototransition and from the Schiff base to Glu-169 during P2 (380) formation. The unusual charge neutrality of both Schiff base counterions in the P2 (380) conducting state suggests that these residues may function as part of a cation selective filter in the open channel state of CaChR1 as well as other low-efficiency ChRs. PMID:25802337

  20. The primary photoreaction of channelrhodopsin-1: Wavelength dependent photoreactions induced by ground-state heterogeneity

    Directory of Open Access Journals (Sweden)

    Karsten eHeyne

    2015-07-01

    Full Text Available The primary photodynamics of channelrhodopsin-1 from Chlamydomonas augustae (CaChR1 was investigated by VIS-pump supercontinuum probe experiments from femtoseconds to 100 picoseconds. In contrast to reported experiments on channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2, we found a clear dependence of the photoreaction dynamics on varying the excitation wavelength. Upon excitation at 500 nm and at 550 nm we detected different bleaching bands, andspectrally distinct photoproduct absorptions in the first picoseconds. We assign the former to the ground-state heterogeneity of a mixture of 13-cis and all-trans retinal maximally absorbing around 480 nm and 540 nm, respectively. At 550 nm, all-trans retinal of the ground state is almost exclusively excited. Here, we found a fast all-trans to 13-cis isomerization process to a hot and spectrally broad P1 photoproduct with a time constant of (100±50 fs, followed by photoproduct relaxation with time constants of (500±100 fs and (5±1 ps. The remaining fraction relaxes back to the parent ground state with time constants of (500±100 fs and (5±1 ps. Upon excitation at 500 nm a mixture of both chromophore conformations is excited, resulting in overlapping reaction dynamics with additional time constants of <300 fs, (1.8±0.3 ps and (90±25 ps. A new photoproduct Q is formed absorbing at around 600 nm. Strong coherent oscillatory signals were found pertaining up to several picoseconds. We determined low frequency modes around 200 cm-1, similar to those reported for bacteriorhodopsin.

  1. A new class of purple membrane variants for the construction of highly oriented membrane assemblies on the basis of noncovalent interactions.

    Science.gov (United States)

    Baumann, Roelf-Peter; Busch, Annegret P; Heidel, Björn; Hampp, Norbert

    2012-04-12

    Purple membranes (PM) from Halobacterium salinarum have been discussed for several technical applications. These ideas started just several years after its discovery. The biological function of bacteriorhodopsin (BR), the only protein in PM, is the light-driven proton translocation across the membrane thereby converting light energy into chemical energy. The astonishing physicochemical robustness of this molecular assembly and the ease of its isolation triggered ideas for technical uses. All basic molecular functions of BR, that is, photochromism, photoelectrism, and proton pumping, are key elements for technical applications like optical data processing and data storage, ultrafast light detection and processing, and direct utilization of sunlight in adenosine 5'-triphospate (ATP) generation or seawater desalination. In spite of the efforts of several research groups worldwide, which confirmed the proof-of-principle for all these potential applications, only the photochromism-based applications have reached a technical level. The physical reason for this is that no fixation or orientation of the PMs is required. The situation is quite different for photoelectrism and proton pumping where the macroscopic orientation of PMs is a prerequisite. For proton pumping, in addition, the formation of artificial membranes which prevent passive proton leakage is necessary. In this manuscript, we describe a new class of PM variants with oppositely charged membrane sides which enable an almost 100% orientation on a surface, which is the key element for photoelectric applications of BR. As an example, the mutated BR, BR-E234R7, was prepared and analyzed. A nearly 100% self-orientation on mica was obtained. PMID:22420766

  2. All-optical light modulation in pharaonis phoborhodopsin and its application to parallel logic gates

    Science.gov (United States)

    Sharma, Parag; Roy, Sukhdev

    2004-08-01

    All-optical light modulation in pharaonis phoborhodopsin (ppR) protein has been analyzed considering its ppRO state dynamics based on nonlinear intensity-induced excited-state absorption. Amplitude modulation of a cw probe laser beam transmission at 560nm corresponding to the peak absorption of ppRO intermediate state through ppR, by a modulating cw pump laser beam at 498nm corresponding to the peak absorption of initial ppR state has been analyzed considering all six intermediate states in its photocylce using the rate equation approach. The transmission characteristics have been shown to exhibit a dip at relatively lower pump intensity values compared to bacteriorhodopsin, which is sensitive to normalized small-signal absorption coefficient (β ), rate constants of ppRM and ppRO states, and absorption of the ppRO state at 498nm. There is an optimum value of β for a given pump intensity range for which maximum modulation can be achieved. It is shown that 100% modulation can be achieved if the initial state of ppR does not absorb the probe beam. The results have been used to design low power all optical parallel NOT, AND, OR, XNOR, and the universal NAND and NOR logic gates for two cases: (i) only changing the output threshold and (ii) considering a common threshold with different β values. At typical parameters, wild-type (WT) ppR based logic gates can be realized at considerably lower pump powers than WT-bR.

  3. Exploring the active site structure of photoreceptor proteins by Raman optical activity

    Science.gov (United States)

    Unno, Masashi

    2015-03-01

    Understanding protein function at the atomic level is a major challenge in a field of biophysics and requires the combined efforts of structural and functional methods. We use photoreceptor proteins as a model system to understand in atomic detail how a chromophore and a protein interact to sense light and send a biological signal. A potential technique for investigating molecular structures is Raman optical activity (ROA), which is a spectroscopic method with a high sensitivity to the structural details of chiral molecules. However, its application to photoreceptor proteins has not been reported. Thus we have constructed ROA spectrometer using near-infrared (NIR) laser excitation at 785 nm. The NIR excitation enables us to measure ROA spectra for a variety of biological samples, including photoreceptor proteins, without fluorescence from the samples. In the present study, we have applied the NIR-ROA to bacteriorhodopsin (BR) and photoactive yellow protein (PYP). BR is a light-driven proton pump and contains a protonated Schiff base of retinal as a chromophore. PYP is a blue light receptor, and this protein has the 4-hydroxycinnamyl chromophore, which is covalently linked to Cys69 through a thiolester bond. We have successfully obtained the ROA spectra of the chromophore within a protein environment. Furthermore, calculations of the ROA spectra utilizing density functional theory provide detailed structural information, such as data on out-of-plane distortions of the chromophore. The structural information obtained from the ROA spectra includes the positions of hydrogen atoms, which are usually not detected in the crystal structures of biological samples.

  4. Photocycle of halorhodopsin from Halobacterium salinarium.

    Science.gov (United States)

    Váró, G; Zimányi, L; Fan, X; Sun, L; Needleman, R; Lanyi, J K

    1995-05-01

    The light-driven chloride pump, halorhodopsin, is a mixture containing all-trans and 13-cis retinal chromophores under both light and dark-adapted conditions and can exist in chloride-free and chloride-binding forms. To describe the photochemical cycle of the all-trans, chloride-binding state that is associated with the transport, and thereby initiate study of the chloride translocation mechanism, one must first dissect the contributions of these species to the measured spectral changes. We resolved the multiple photochemical reactions by determining flash-induced difference spectra and photocycle kinetics in halorhodopsin-containing membranes prepared from Halobacterium salinarium, with light- and dark-adapted samples at various chloride concentrations. The high expression of cloned halorhodopsin made it possible to do these measurements with unfractionated cell envelope membranes in which the chromophore is photostable not only in the presence of NaCl but also in the Na2SO4 solution used for reference. Careful examination of the flash-induced changes at selected wavelengths allowed separating the spectral changes into components and assigning them to the individual photocycles. According to the results, a substantial revision of the photocycle model for H. salinarium halorhodopsin, and its dependence on chloride, is required. The cycle of the all-trans chloride-binding form is described by the scheme, HR-hv-->KL1L2N-->HR, where HR, K, L, and N designate halorhodopsin and its photointermediates. Unlike the earlier models, this is very similar to the photoreaction of bacteriorhodopsin when deprotonation of the Schiff base is prevented (e.g., at low pH or in the D85N mutant). Also unlike in the earlier models, no step in this photocycle was noticeably affected when the chloride concentration was varied between 20 mM and 2 M in an attempt to identify a chloride-binding reaction. PMID:7612849

  5. Microarray analysis in the archaeon Halobacterium salinarum strain R1.

    Directory of Open Access Journals (Sweden)

    Jens Twellmeyer

    Full Text Available BACKGROUND: Phototrophy of the extremely halophilic archaeon Halobacterium salinarum was explored for decades. The research was mainly focused on the expression of bacteriorhodopsin and its functional properties. In contrast, less is known about genome wide transcriptional changes and their impact on the physiological adaptation to phototrophy. The tool of choice to record transcriptional profiles is the DNA microarray technique. However, the technique is still rarely used for transcriptome analysis in archaea. METHODOLOGY/PRINCIPAL FINDINGS: We developed a whole-genome DNA microarray based on our sequence data of the Hbt. salinarum strain R1 genome. The potential of our tool is exemplified by the comparison of cells growing under aerobic and phototrophic conditions, respectively. We processed the raw fluorescence data by several stringent filtering steps and a subsequent MAANOVA analysis. The study revealed a lot of transcriptional differences between the two cell states. We found that the transcriptional changes were relatively weak, though significant. Finally, the DNA microarray data were independently verified by a real-time PCR analysis. CONCLUSION/SIGNIFICANCE: This is the first DNA microarray analysis of Hbt. salinarum cells that were actually grown under phototrophic conditions. By comparing the transcriptomics data with current knowledge we could show that our DNA microarray tool is well applicable for transcriptome analysis in the extremely halophilic archaeon Hbt. salinarum. The reliability of our tool is based on both the high-quality array of DNA probes and the stringent data handling including MAANOVA analysis. Among the regulated genes more than 50% had unknown functions. This underlines the fact that haloarchaeal phototrophy is still far away from being completely understood. Hence, the data recorded in this study will be subject to future systems biology analysis.

  6. Proteins as "dopable" bio-electronic materials

    Science.gov (United States)

    Cahen, David

    2013-02-01

    Proteins are surprisingly good solid-state electronic conductors. This holds also for proteins without any known biological electron transfer function. How do they do it? To answer this question we measure solid-state electron transport (ETp) across proteins that are "dry" (only tightly bound water, to retain the conformation, still present). We compare results for the electron transfer (ET) protein, Azurin (Az), the proton-pumping membrane protein Bacteriorhodopsin (bR), and for Human and Bovine Serum Albumin (HSA and BSA). Clear differences between these proteins are seen, which preserve their structure in the solid state measurement configuration. Importantly for future bioelectronics, the results are sensitive to protein modification, e.g., removing or disconnecting the retinal in bR and removing or replacing the Cu redox centre in Az. These cofactors can thus be viewed as natural dopants for proteins. Insight in the ETp mechanism comes from temperature-dependent studies. Az shows 40-360K temperature-independent ETp across its 3.5 nm long axis, until its denaturation temperature, indicative of tunneling. Cu removal, replacement (by Zn) or deuteration changes this to thermally activated ETp. This suggests hopping and involvement of the amide backbone in the ETp. The latter, which rhymes with indications from ETp experiments on oligopeptide and simulations of ET in proteins, opens the way for modeling what otherwise is an awfully complex system. Below 200K all proteins and their variants show temperature-independent ETp. We can furthermore make a totally electrically inactive protein, HSA, into an efficient ETp medium by doping it with natural poly-ene. Putting our data in perspective by comparing them to all known protein ETp data in the literature, we conclude that, in general, proteins are well described as dopable molecular wires.

  7. Light-induced membrane potential and pH gradient in Halobacterium halobium envelope vesicles.

    Science.gov (United States)

    Renthal, R; Lanyi, J K

    1976-05-18

    Illumination of envelope vesicles prepared from Halobacterium halobium cells causes translocation of protons from inside to outside, due to the light-induced cycling of bacteriorhodopsin. This process results in a pH gradient across the membranes, an electrical potential, and the movements of K+ and Na+. The electrical potential was estimated by following the fluorescence of a cyanine dye, 3,3'-dipentyloxadicarbocyanine. Illumination of H. halobium vesicles resulted in a rapid, reversible decrease of the dye fluorescence, by as much as 35%. This effect was not seen in nonvesicular patches of purple membrane. Observation of maximal fluorescence decreases upon ilumination of vesicles required an optimal dye/membrane protein ratio. The pH optimum for the lightinduced fluorescence decrease was 6.0. The decrease was linear with actinic light intensity up to about 4 X 10(5) ergs cn-2 s-1. Valinomycin, gramicidin, and triphenylmethylphosphonium ion all abolished the fluorescence changes. However, the light-induced pH change was enhanced by these agents. Conversely, buffered vesicles showed no pH change but gave the same or larger fluorescence changes. Thus, we have identified the fluorescence decrease with a light-induced membrane potential, inside negative. By using valinomycin-K+-induced membrane potentials, we calibrated the fluorescence decrease with calculated Nernst diffusion potentials. We found a linear dependence between potential and fluorescence decrease of 3 mV/%, up to 90 mV. When the envelope vesicles were illuminated, the total proton-motive force generated was dependent on the presence of Na+ and K+ and their concentration gradients across the membrane. In general, K+ appeared to be more permeable than Na+ and, thus, permitted development of greater pH gradients and lower electrical potentials. By calculating the total proton-motive force from the sum of the pH and potential terms, we found that the vesicles can produce proton-motive forces near--200 m

  8. F-state based p ermanent optical data storage in bacteriorho dopsin film%基于菌紫质F态的永久光存储研究∗

    Institute of Scientific and Technical Information of China (English)

    于湘华; 但旦; 闵俊伟; 严绍辉; 姚保利; 李新宇; 刘石磊; 雷铭; 李润泽; 梁言生; 周兴; 吴迪

    2015-01-01

    在线偏振飞秒激光激发下,菌紫质通过双光子光化学反应可以生成具有永久光致各向异性的蓝移产物F540态。基于F540态的永久光致各向异性,通过调控飞秒激光空间光场分布,可以在菌紫质薄膜中实现永久光信息存储。本文使用纯相位型空间光调制器调制飞秒激光光场,在物镜焦平面上生成光学点阵图案,可以将信息快速记录在菌紫质薄膜中。同时,通过改变入射激光偏振方向,可以实现偏振复用光存储,这在高密度光存储和数据加密领域具有潜在应用。%Upon excitation of high intensity linearly polarized femtosecond laser, a blue-shifted two-photon induced photo-product named F540 state with permanent photo-induced anisotropy is observed in bacteriorhodopsin (BR) films. Based on the F-state, permanent optical data storage can be realized with spatial field modulation of femtosecond laser. By using a phase-only spatial light modulator to modulate the spatial phase distribution of the incident femtosecond laser beam, we generate spot-patterns in the focal plane of the microscopic objective lens, and the patterns are recorded in the BR film. At the same time, by varying the polarization direction of the excitation laser beam, polarization-multiplexed optical data storage in the BR film is demonstrated, which is applicable to high-density optical data storage and optical information encryption.

  9. Comparison of the structural changes occurring during the primary phototransition of two different channelrhodopsins from Chlamydomonas algae.

    Science.gov (United States)

    Ogren, John I; Yi, Adrian; Mamaev, Sergey; Li, Hai; Lugtenburg, Johan; DeGrip, Willem J; Spudich, John L; Rothschild, Kenneth J

    2015-01-20

    Channelrhodopsins (ChRs) from green flagellate algae function as light-gated ion channels when expressed heterologously in mammalian cells. Considerable interest has focused on understanding the molecular mechanisms of ChRs to bioengineer their properties for specific optogenetic applications such as elucidating the function of specific neurons in brain circuits. While most studies have used channelrhodopsin-2 from Chlamydomonas reinhardtii (CrChR2), in this work low-temperature Fourier transform infrared-difference spectroscopy is applied to study the conformational changes occurring during the primary phototransition of the red-shifted ChR1 from Chlamydomonas augustae (CaChR1). Substitution with isotope-labeled retinals or the retinal analogue A2, site-directed mutagenesis, hydrogen-deuterium exchange, and H2(18)O exchange were used to assign bands to the retinal chromophore, protein, and internal water molecules. The primary phototransition of CaChR1 at 80 K involves, in contrast to that of CrChR2, almost exclusively an all-trans to 13-cis isomerization of the retinal chromophore, as in the primary phototransition of bacteriorhodopsin (BR). In addition, significant differences are found for structural changes of the protein and internal water(s) compared to those of CrChR2, including the response of several Asp/Glu residues to retinal isomerization. A negative amide II band is identified in the retinal ethylenic stretch region of CaChR1, which reflects along with amide I bands alterations in protein backbone structure early in the photocycle. A decrease in the hydrogen bond strength of a weakly hydrogen bonded internal water is detected in both CaChR1 and CrChR2, but the bands are much broader in CrChR2, indicating a more heterogeneous environment. Mutations involving residues Glu169 and Asp299 (homologues of the Asp85 and Asp212 Schiff base counterions, respectively, in BR) lead to the conclusion that Asp299 is protonated during P1 formation and suggest that

  10. Retinal chromophore structure and Schiff base interactions in red-shifted channelrhodopsin-1 from Chlamydomonas augustae.

    Science.gov (United States)

    Ogren, John I; Mamaev, Sergey; Russano, Daniel; Li, Hai; Spudich, John L; Rothschild, Kenneth J

    2014-06-24

    Channelrhodopsins (ChRs), which form a distinct branch of the microbial rhodopsin family, control phototaxis in green algae. Because ChRs can be expressed and function in neuronal membranes as light-gated cation channels, they have rapidly become an important optogenetic tool in neurobiology. While channelrhodopsin-2 from the unicellular alga Chlamydomonas reinhardtii (CrChR2) is the most commonly used and extensively studied optogenetic ChR, little is known about the properties of the diverse group of other ChRs. In this study, near-infrared confocal resonance Raman spectroscopy along with hydrogen-deuterium exchange and site-directed mutagenesis were used to study the structure of red-shifted ChR1 from Chlamydomonas augustae (CaChR1). These measurements reveal that (i) CaChR1 has an all-trans-retinal structure similar to those of the light-driven proton pump bacteriorhodopsin (BR) and sensory rhodopsin II but different from that of the mixed retinal composition of CrChR2, (ii) lowering the pH from 7 to 2 or substituting neutral residues for Glu169 or Asp299 does not significantly shift the ethylenic stretch frequency more than 1-2 cm(-1) in contrast to BR in which a downshift of 7-9 cm(-1) occurs reflecting neutralization of the Asp85 counterion, and (iii) the CaChR1 protonated Schiff base (SB) has stronger hydrogen bonding than BR. A model is proposed to explain these results whereby at pH 7 the predominant counterion to the SB is Asp299 (the homologue to Asp212 in BR) while Glu169 (the homologue to Asp85 in BR) exists in a neutral state. We observe an unusual constancy of the resonance Raman spectra over the broad range from pH 9 to 2 and discuss its implications. These results are in accord with recent visible absorption and current measurements of CaChR1 [Sineshchekov, O. A., et al. (2013) Intramolecular proton transfer in channelrhodopsins. Biophys. J. 104, 807-817; Li, H., et al. (2014) Role of a helix B lysine residue in the photoactive site in

  11. Alpha-helical hydrophobic polypeptides form proton-selective channels in lipid bilayers

    Science.gov (United States)

    Oliver, A. E.; Deamer, D. W.

    1994-01-01

    Proton translocation is important in membrane-mediated processes such as ATP-dependent proton pumps, ATP synthesis, bacteriorhodopsin, and cytochrome oxidase function. The fundamental mechanism, however, is poorly understood. To test the theoretical possibility that bundles of hydrophobic alpha-helices could provide a low energy pathway for ion translocation through the lipid bilayer, polyamino acids were incorporated into extruded liposomes and planar lipid membranes, and proton translocation was measured. Liposomes with incorporated long-chain poly-L-alanine or poly-L-leucine were found to have proton permeability coefficients 5 to 7 times greater than control liposomes, whereas short-chain polyamino acids had relatively little effect. Potassium permeability was not increased markedly by any of the polyamino acids tested. Analytical thin layer chromatography measurements of lipid content and a fluorescamine assay for amino acids showed that there were approximately 135 polyleucine or 65 polyalanine molecules associated with each liposome. Fourier transform infrared spectroscopy indicated that a major fraction of the long-chain hydrophobic peptides existed in an alpha-helical conformation. Single-channel recording in both 0.1 N HCl and 0.1 M KCl was also used to determine whether proton-conducting channels formed in planar lipid membranes (phosphatidylcholine/phosphatidylethanolamine, 1:1). Poly-L-leucine and poly-L-alanine in HCl caused a 10- to 30-fold increase in frequency of conductive events compared to that seen in KCl or by the other polyamino acids in either solution. This finding correlates well with the liposome observations in which these two polyamino acids caused the largest increase in membrane proton permeability but had little effect on potassium permeability. Poly-L-leucine was considerably more conductive than poly-L-alanine due primarily to larger event amplitudes and, to a lesser extent, a higher event frequency. Poly-L-leucine caused two

  12. Systematic analysis of protein–detergent complexes applying dynamic light scattering to optimize solutions for crystallization trials

    International Nuclear Information System (INIS)

    Application of in situ dynamic light scattering to solutions of protein–detergent complexes permits characterization of these complexes in samples as small as 2 µl in volume. Detergents are widely used for the isolation and solubilization of membrane proteins to support crystallization and structure determination. Detergents are amphiphilic molecules that form micelles once the characteristic critical micelle concentration (CMC) is achieved and can solubilize membrane proteins by the formation of micelles around them. The results are presented of a study of micelle formation observed by in situ dynamic light-scattering (DLS) analyses performed on selected detergent solutions using a newly designed advanced hardware device. DLS was initially applied in situ to detergent samples with a total volume of approximately 2 µl. When measured with DLS, pure detergents show a monodisperse radial distribution in water at concentrations exceeding the CMC. A series of all-transn-alkyl-β-d-maltopyranosides, from n-hexyl to n-tetradecyl, were used in the investigations. The results obtained verify that the application of DLS in situ is capable of distinguishing differences in the hydrodynamic radii of micelles formed by detergents differing in length by only a single CH2 group in their aliphatic tails. Subsequently, DLS was applied to investigate the distribution of hydrodynamic radii of membrane proteins and selected water-insoluble proteins in presence of detergent micelles. The results confirm that stable protein–detergent complexes were prepared for (i) bacteriorhodopsin and (ii) FetA in complex with a ligand as examples of transmembrane proteins. A fusion of maltose-binding protein and the Duck hepatitis B virus X protein was added to this investigation as an example of a non-membrane-associated protein with low water solubility. The increased solubility of this protein in the presence of detergent could be monitored, as well as the progress of proteolytic cleavage to

  13. Systematic analysis of protein–detergent complexes applying dynamic light scattering to optimize solutions for crystallization trials

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, Arne [University of Hamburg, c/o DESY, Building 22a, Notkestrasse 85, 22603 Hamburg (Germany); Dierks, Karsten [University of Hamburg, c/o DESY, Building 22a, Notkestrasse 85, 22603 Hamburg (Germany); XtalConcepts, Marlowring 19, 22525 Hamburg (Germany); Hussein, Rana [University of Hamburg, c/o DESY, Building 22a, Notkestrasse 85, 22603 Hamburg (Germany); Brillet, Karl [ESBS, Pôle API, 300 Boulevard Sébastien Brant, CS10413, 67412 Illkirch CEDEX (France); Brognaro, Hevila [São Paulo State University, UNESP/IBILCE, Caixa Postal 136, São José do Rio Preto-SP, 15054 (Brazil); Betzel, Christian, E-mail: christian.betzel@uni-hamburg.de [University of Hamburg, c/o DESY, Building 22a, Notkestrasse 85, 22603 Hamburg (Germany)

    2015-01-01

    Application of in situ dynamic light scattering to solutions of protein–detergent complexes permits characterization of these complexes in samples as small as 2 µl in volume. Detergents are widely used for the isolation and solubilization of membrane proteins to support crystallization and structure determination. Detergents are amphiphilic molecules that form micelles once the characteristic critical micelle concentration (CMC) is achieved and can solubilize membrane proteins by the formation of micelles around them. The results are presented of a study of micelle formation observed by in situ dynamic light-scattering (DLS) analyses performed on selected detergent solutions using a newly designed advanced hardware device. DLS was initially applied in situ to detergent samples with a total volume of approximately 2 µl. When measured with DLS, pure detergents show a monodisperse radial distribution in water at concentrations exceeding the CMC. A series of all-transn-alkyl-β-d-maltopyranosides, from n-hexyl to n-tetradecyl, were used in the investigations. The results obtained verify that the application of DLS in situ is capable of distinguishing differences in the hydrodynamic radii of micelles formed by detergents differing in length by only a single CH{sub 2} group in their aliphatic tails. Subsequently, DLS was applied to investigate the distribution of hydrodynamic radii of membrane proteins and selected water-insoluble proteins in presence of detergent micelles. The results confirm that stable protein–detergent complexes were prepared for (i) bacteriorhodopsin and (ii) FetA in complex with a ligand as examples of transmembrane proteins. A fusion of maltose-binding protein and the Duck hepatitis B virus X protein was added to this investigation as an example of a non-membrane-associated protein with low water solubility. The increased solubility of this protein in the presence of detergent could be monitored, as well as the progress of proteolytic

  14. Models of the Protocellular Structures, Functions and Evolution

    Science.gov (United States)

    Pohorille, Andrew; New, Michael; Keefe, Anthony; Szostak, Jack W.; Lanyi, Janos F.; DeVincenzi, Donald L. (Technical Monitor)

    2000-01-01

    environment. To provide a continuous energy supply, usually needed to activate the substrates, an energy transduction complex which generates ATP from adenosine diphosphate, inorganic phosphate and light will be used. This system, consisting of two modern proteins, ATP synthase and bacteriorhodopsin, has already been built and shown to work efficiently. By coupling chemical synthesis to such a system, it will be possible to drive chemical reactions by light if only the substrates for these reactions are supplied.

  15. Ion-beam and microwave-stimulated functionalization and derivatization of carbon nanotubes

    Science.gov (United States)

    Makala, Raghuveer S.

    Derivatizing carbon nanotubes (CNTs) with other low-dimensional nanostructures is of widespread interest for creating CNT-based nanocomposites and devices. Conventional routes based on wet-chemical oxidation or hydrophobic adsorption do not allow premeditated control over the location or spatial extent of functionalization. Moreover, aggressive oxidative treatments and agitation in corrosive environments lead to CNT shortening, damage, and incorporation of excess impurity concentrations. Thus, it is imperative to explore and develop alternative functionalization methods to overcome these shortcomings. The work presented in this thesis outlines two such methodologies: one based on focused ion irradiation for siteselective functionalization and the other that employs microwave-stimulation for mild, yet rapid and homogenous CNT functionalization. The utility of 10 and 30 kcV Ga+ focused ion beams (FIB) to thin, slice, weld, and alter the structure and composition at precise locations along the CNT axis is presented. This strategy of harnessing ion-beam-induced defect generation and doping is attractive for modulating chemical and electrical properties along the CNT length, and fabricate CNT-based heterostructures and networks. A novel approach that utilizes focused ion irradiation to site-selectively derivatize preselected segments of CNTs with controlled micro-/nano-scale lateral spatial resolution is demonstrated. Irradiation followed by air-exposure results in functionalized CNT segments ranging from the nanoscopic to the macroscopic scale. The functional moieties are utilized to site-selectively anchor Au nanoparticles, fluorescent nanospheres, an amino acid---lysine, a charge-transfer metalloprotein---azurin, and a photoactive protein---bacteriorhodopsin by means of electrostatic or covalent interactions. This approach is versatile and can be extended to obtaining other molecular moieties and derivatives opening up possibilities for building new types of nano

  16. Energy harvesting with bio-inspired synthetic nanochannels%基于仿生智能纳米孔道的先进能源转换体系

    Institute of Scientific and Technical Information of China (English)

    郭维; 江雷

    2011-01-01

    With more than four billion years' evolution and selection, natural creatures inherit almost perfect functional structures to harvesting clean energy from their living environments. Particularly, biological ion channels and pumps on cell membrane play a crucial role in many bioelectrogenesis processes. Learning from Nature, such as the electric eels, the synthesis of ATP molecules, the Bacteriorhodopsin on retina, etc, provides new approaches for the construction of novel energy conversion systems. In this review article, we briefly summarize the three most intensively studied topics on the clean energy conversion system with bio-inspired nanochannels and nanopores, which are the nanofluidic electrokinetic conversion systems, the nanofluidic reverse electrodialysis systems, and these newly arisen advanced energy conversion system with smart synthetic ion channels. In a predictable future, the performance of these bio-inspired energy conversion system will surly exceed the present achievements of the conventional man-made systems. The bio-mimetic and bio-inspired strategies provide new insights into future energy conversion techniques.%自然界中的生命体系经过40多亿年的进化,实现了对能源的高效转换、存储和利用.特别是生物膜上的各类孔道结构在其中起着重要作用.基于仿生智能纳米通道的先进能源转换体系从生物离子通道中获取与能量转换相关的启示(例如,电鳗放电、ATP合成、视网膜、紫膜等),从原理和结构上模仿生命体系中高效能量转换的某一个侧面,通过产能材料的设计和转换器件的组装,实现机械能到电能、光能到电能、光能到化学能等不同能量形式之间的转换.我们综述了目前应用人工合成的纳米尺度孔道结构进行仿生能源转换的三个热点领域:纳米流体动能-电能转换,纳米流体反向电渗析系统和基于仿生智能纳米孔道的先进能源转换体系.基于智能纳米孔道

  17. Solar Energy Program: Chapter from the Energy and EnvironmentalDivision Annual Report 1980

    Energy Technology Data Exchange (ETDEWEB)

    Energy and Environment Division

    1981-03-01

    -term possibilities for utilizing solar energy. One project involves the development of a new type of solar thermal receiver that would be placed at the focus of a central receiver system or a parabolic dish. The conversion of the concentrated sunlight to thermal energy would be accomplished by the absorption of the light by a dispersion of very small particles suspended in a gas. Another project is exploring biological systems. In particular, we are investigating the possibility of developing a photovoltaic cell, based on a catalyst (bacteriorhodopsin) which converts light to electrical ion flow across the cell membrane of a particular bacteria.

  18. Enhanced Oil Recovery by Horizontal Waterflooding

    Energy Technology Data Exchange (ETDEWEB)

    Scott Robinowitz; Dwight Dauben; June Schmeling

    2005-09-05

    -term possibilities for utilizing solar energy. One project involves the development of a new type of solar thermal receiver that would be placed at the focus of a central receiver system or a parabolic dish. The conversion of the concentrated sunlight to thermal energy would be accomplished by the absorption of the light by a dispersion of very small particles suspended in a gas. Another project is exploring biological systems. In particular, we are investigating the possibility of developing a photovoltaic cell, based on a catalyst (bacteriorhodopsin) which converts light to electrical ion flow across the cell membrane of a particular bacteria.

  19. The extreme environments and their microbes as models for extraterrestrial life

    Science.gov (United States)

    Seckbach, J.; Oren, A.; Chela-Flores, J.

    2008-09-01

    Bacteria such as the aerobic Salinibacter ruber and the anaerobic members of the Halanaerobiales) use KCl to provide the necessary osmotic balance. Some of these extreme halophiles possess light-driven proton pumps (bacteriorhodopsin, xanthorhodopsin) and chloride pumps (halorhodopsin) that enable them to use photons to drive energetically expensive reactions (Oren, 2002; Oren, 2008). Extremophiles can serve as models for extraterrestrial microbes that may live in celestial bodies. The most promising among these to contain habitable areas are Mars (where the Phoenix Lander recently discovered water) and the Jovian satellite Europa; also Titan (the moon of Saturn) has some features that resemble those that may have existed on Earth during its earliest stages. From the characteristics of extremophilic microorganisms found on the present-day Earth, we can derive some insights on the question of habitability of other planets, and learn about possible bioindicators that may be suitable when searching for extraterrestrial life (Seckbach and Chela-Flores, 2007). Compounds such as methane on Mars or traces of sulfur on Jupiter's moon Europa may have been of biogenic origin and may possibly have been endogenic (Chela-Flores, 2006; Chela-Flores and Kumar, 2008). Biogeochemical tests have been proposed for missions that are in the planning stages, such as LAPLACE (Blanc et al., 2008), a mission to Europa and the Jupiter system by ESA's Cosmic Vision Programme. The finding of elemental sulfur on Europa may be of special interest. One possibility is that such traces of sulfur might have originated from the metabolism of extremophilic sulfurreducing microorganisms. Radiation may damage traces of biogenic sulfur deposited on the surface. The stopping depth for ionic radiation in the Jovian magnetosphere is expected not to exceed 1 cm (Greenberg, 2005; Dudeja et al., 2008). Thus, organic molecules would not be destroyed below such a thin layer. Based on to the preliminary results of the