电极介导的菌紫质干膜微分光电流响应的整流性质

在长方形光脉冲光照下,菌紫质(bacteriorhodopsin,BR)干膜组装成夹层光电池具有微分光电流响应．在氧化铟锡(ITO)导电玻璃/BR膜/封口膜/不锈钢形成的干膜电池下可观察到整流特性,而在不锈钢/BR膜/封口膜/ITO导电玻璃形成的干膜电池下则观察不到整流特性,这说明是电极介导的整流．平衡电压测定表明：工作电极/BR膜表面与对电极/BR膜表面有不同的性质,电极的界面效应控制了BR的取向．酸与碱产生的瞬间电流极性也证实了电极整流行为的存在．这些结果将有助于了解BR膜的微分光电响应．     

细菌视紫红质的光电响应特性和机制

在ITO导电玻璃上制备定向细菌视紫红质 (BR)电泳沉积膜或LB膜组成光电池系统 ,在短脉冲激光照射下 ,测定其脉冲响应光电压 ;在间断光照射下 ,测定其对光强变化产生的微分响应信号。对脉冲光电响应和微分响应的机理及其关系进行理论分析和解释 ,认为脉冲响应是BR分子内部生色团快速光极化引起的电荷分离和希夫碱及其周围氨基酸去质子化和再质子化过程引起的质子定向运输产生的位移电流 ,是一个快反应过程 ,是微分响应的早期反应和基础。微分响应则是由于菌紫质的光驱动质子泵产生的连续质子流在光开和光关瞬间引起光电池系统充放电以及测量电路的耦合特性引起的 ,是一个慢变化过程     

温度和pH对嗜盐菌紫膜吸收和圆二色谱的作用

嗜盐菌紫膜可见光区域的圓二色谱由二部分重迭组成,一部分是较宽的正带,它来自细菌视紫红质分子内视黄醛和菌蛋白之间相互作用,另一部分是由对称的正带、负带组成的激子带,它来自膜内相邻细菌视紫红质分子生色团的相互作用。因此,紫膜的圓二色谱可反映膜上蛋白的晶体结构和构象变化,被认为是研究嗜盐菌紫膜结构变化的一种天然探针。本文用圆二色谱和吸收谱观察了温度和pH对光适应紫膜结构的影响。     

细菌视紫红质光学图像存储的灰阶特性研究

细菌视紫红质（Bacteriorhodopsin,BR）是一种具有优良光致变色特性的光敏蛋白分子,具有极好的抗疲劳性和高的光转换量子效率,可用于光学图像信息的获取和存储.文章讨论了BR分子膜在受到随空间位置变化的光强调制下,BR分子膜光吸收变化量的空间分布及其与存储图像灰度分布之间的关系;建立了BR灰度图像存储特性实验系统,并对BR-D96N薄膜存储的光学图像灰阶特性进行了实验研究,实验表明BR薄膜图像存储具有出色的灰度表现能力.     

细菌视紫红质的两种光电微分响应及其机制

利用MATLAB软件对细菌视紫红质 (BR)膜光电器件的脉冲响应实验数据进行拟合 ,得到器件的冲激响应函数。据此用SIMULINK模块构造出了反映BR光电器件特性的仿真系统。利用此系统对不同间断光入射BR光电器件时的输出响应信号进行了仿真计算。通过分析得出结论 :以前所描述的微分响应 (发生在毫秒到秒的时间量级 ,在光打开时产生一个正脉冲 ,在光关掉时产生一个负脉冲 )并非BR分子固有的特性 ,部分是由于测量电路引起的。BR分子本身特性引起的微分响应是发生在微秒时间量级 ,而且在光打开时产生一个负脉冲 ,在光关掉时产生一个正脉冲。对这两种微分响应产生的机制分别进行了探讨     

高碱性pH诱导紫膜片层表面结构变化的原子力显微镜直接观测

报道了在高碱性pH下,紫膜中细菌视紫红质(BR)表面结构变化的直观信息.紫外可见光谱实验发现,当pH上升到12.6,BR分子上的生色团视黄醛脱落,分子完全变性;原子力显微镜实验观测到在此pH下,紫膜片层的晶格结构瓦解,BR分子在紫膜上无规则聚集,同时出现非特征“岛屿”结构和特征“岛屿”结构.     

细菌视紫红质中质子泵出的分子机理

细菌视紫红质(Bacteriorhodopsin,或bR)是盐生嗜盐菌(Halobacterium salinarium)等细菌的跨膜蛋白质,其色基视黄醛的光致异构化作用触发细菌视紫红质的一系列结构变化,把质子从细胞质泵到细胞外空间。对细菌视紫红质中质子泵出分子机理进行了描述。     

菌紫质人工膜的光电压与离子强度的非线性关系

本文研究了Ｋ＋、Ｍｇ２＋和Ｌａ３＋离子强度对菌紫质人工膜的光电压的影响。研究结果表明,在实验所测定的离子强度范围内菌紫质人工沉淀膜的光电压与上述离子的离子强度间具有非线性关系,并随离子价数的提高其响应峰值的位置向低浓度发展。本文研究结果将对菌紫质光化学反应循环、质子迁移机制的研究,以及菌紫质光电池的设计、和以菌紫质为基础的视觉功能模拟和人工视觉研究提供有价值的数据 。     

菌紫质人工膜的光电压与离子强度的非线性关系

本文研究了Ｋ＾＋,Ｍｇ＾２＋和Ｌａ＾３＋离子强度对菌紫质人工膜的光电压的影响。研究结果表明,在实验所测定的离子强度范围的内菌紫质人工沉淀膜的光电压与上述离子的离子强度间具有非线性关系,并随离子价数的提高其响应峰值的位置向低浓度发展。本文研究结果将对菌紫紫质光化学反应循环,质子迁移机制的研究,以及菌紫质光电池的设计,和以菌紫质为基础的视觉功能模拟和人工视觉研究提供有价值的数据。     

紫膜菌紫质分子的光生物物理学研究

本文介绍了紫膜细菌视紫红质的微观结构,质子泵功能和光循环过程,及其菌紫质分子中发色团和蛋白质相互作用(结合位点)的几种光谱研究,最后讨论了菌紫质分子原初事件量子产率的双重性和相应的分子动力学模型。     

Light adaptation in Drosophila photoreceptors: I. Response dynamics and signaling efficiency at 25 degrees C

Besides the physical limits imposed on photon absorption, the coprocessing of visual information by the phototransduction cascade and photoreceptor membrane determines the fidelity of photoreceptor signaling. We investigated the response dynamics and signaling efficiency of Drosophila photoreceptors to natural-like fluctuating light contrast stimulation and intracellular current injection when the cells were adapted over a 4-log unit light intensity range at 25 degrees C. This dual stimulation allowed us to characterize how an increase in the mean light intensity causes the phototransduction cascade and photoreceptor membrane to produce larger, faster and increasingly accurate voltage responses to a given contrast. Using signal and noise analysis, this appears to be associated with an increased summation of smaller and faster elementary responses (i.e., bumps), whose latency distribution stays relatively unchanged at different mean light intensity levels. As the phototransduction cascade increases, the size and speed of the signals (light current) at higher adapting backgrounds and, in conjunction with the photoreceptor membrane, reduces the light-induced voltage noise, and the photoreceptor signal-to-noise ratio improves and extends to a higher bandwidth. Because the voltage responses to light contrasts are much slower than those evoked by current injection, the photoreceptor membrane does not limit the speed of the phototransduction cascade, but it does filter the associated high frequency noise. The photoreceptor information capacity increases with light adaptation and starts to saturate at approximately 200 bits/s as the speed of the chemical reactions inside a fixed number of transduction units, possibly microvilli, is approaching its maximum.     

Diurnal modulation of photoreceptor potassium conductance in the locust

Single electrode clamp techniques demonstrated diurnal changes in photoreceptor membrane conductance, recorded intracellularly in the intact, dark-adapted retina of the locust Schistocerca gregaria. In the day, locust photoreceptors exhibited the membrane properties of fast cells, as previously defined in rapidly moving diurnal Diptera. Depolarization activated a powerful potassium conductance with two kinetic components, one rapidly activating close to resting potential and the other activating more slowly when further depolarized, giving a pronounced delayed rectification. There was little inactivation. At night, locust photoreceptors resembled slow cells, as defined in weakly flying crepuscular and nocturnal Diptera. Depolarization rapidly activated an outward current which then inactivated over 100 ms to reduce rectification. The change from day to night state was mimicked by applying 10 mM serotonin extracellularly to the retina. We conclude that the potassium conductances of locust photoreceptor membranes are modulated according to a diurnal rhythm, possibly by serotonin. This neuromodulation is used to match photoreceptor membrane properties to photic habitat. Our findings suggest a definite and potentially widespread function for serotonin as a mediator of diurnal changes in the insect visual system.     

Rhodopsin Receptors of Phototaxis in Green Flagellate Algae

Green flagellate algae are capable of the active adjustment of their swimming path according to the light direction (phototaxis). This direction is detected by a special photoreceptor apparatus consisting of the photoreceptor membrane and eyespot. Receptor photoexcitation in green flagellates triggers a cascade of rapid electrical events in the cell membrane which plays a crucial role in the signal transduction chain of phototaxis and the photophobic response. The photoreceptor current is the earliest so far detectable process in this cascade. Measurement of the photoreceptor current is at present the most suitable approach to investigation of the photoreceptor pigment in green flagellate algae, since a low receptor concentration in the cell makes application of optical and biochemical methods so far impossible. A set of physiological evidences shows that the phototaxis receptor in green flagellate algae is a unique rhodopsin-type protein. It shares common chromophore properties with retinal proteins from archaea. However, the involvement of photoelectric processes in the signal transduction chain relates it to animal visual rhodopsins. The presence of some enzymatic components of the animal visual cascade in isolated eyespot preparations might also point to this relation. A retinal-binding protein has been identified in such preparations, the amino acid sequence of which shows a certain homology to sequences of animal visual rhodopsins. However, potential function of this protein as the phototaxis receptor has been questioned in recent time.     

A model of photoreceptor cell for response to light through synaptic connection

A model is proposed for the responses of vertebrate photoreceptor cell to light stimuli. It is based on the findings that the resistance of visual cell membrane increases during illumination. In this model the relation between the changes of membrane resistance and light intensity through synaptic connection is considered. This model suggests the general relation between the peak amplitude of receptor response and the intensity of flash.     

Membrane filtering properties of the bumblebee (Bombus terrestris) photoreceptors across three spectral classes

Filtering properties of the membrane form an integral part of the mechanisms producing the light-induced electrical signal in insect photoreceptors. Insect photoreceptors vary in response speed between different species, but recently it has also been shown that different spectral photoreceptor classes within a species possess diverse response characteristics. However, it has not been quantified what roles phototransduction and membrane properties play in such diversity. Here, we use electrophysiological methods in combination with system analysis to study whether the membrane properties could create the variation of the response speed found in the bumblebee (Bombus terrestris) photoreceptors. We recorded intracellular responses from each photoreceptor class to white noise-modulated current stimuli and defined their input resistance and linear filtering properties. We found that green sensitive cells exhibit smaller input resistance and membrane impedance than other cell classes. Since green sensitive cells are the fastest photoreceptor class in the bumblebee retina, our results suggest that the membrane filtering properties are correlated with the speed of light responses across the spectral classes. In general, our results provide a compelling example of filtering at the sensory cell level where the biophysical properties of the membrane are matched to the performance requirements set by visual ecology.     

Improved isolation procedures for the purple membrane of Halobacterium halobium.

Techniques for purifying teh purple membrane of Halobacterium halobium are given. This purple membrane contains a chromoprotein with a retinal prosthetic group similar to rhodopsin, the chromprotein found in the visual systems of higher invertebrates and vertebrates. The described purple membrane isolation procedures yield a highly purified preparation as determined by transmitting electron microscopy and gel electrophoresis. Critical analysis of the absorption spectra of the purple membrane was also employed to establish criteria of purity for the preparation. The visible absorption spectra of the purified purple membrane preparation in buffer was found to have a maximum at 559 nm which shifted to 567 nm on light exposure. No indication of any spectral perturbation arising from bacterioruberin-containing membrane, the major contaminant in purple membrane preparations, was found. Furthermore, the ratio of protein aromatic amino acid absorbance at 280 nm to chromophore absorbance at 567 nm was found to be 1.5 in light-exposed preparations compared to the previously reported ratio of 2.3.-3 The decrease in the value of this ratio is also indicative of an increase in the purity of the purple membrane preparation.     

Color image detection by biomolecular photoreceptor using bacteriorhodopsin-based complex LB films

A biomolecular photoreceptor consisting of bacteriorhodopsin (bR)-based complex Langmuir–Blodgett (LB) films was developed for color image detection. By mimicking the functions of the pigments in retina of human visual system, biomolecules with photoelectric conversion function were chosen and used as constituents for an artificial photoreceptor. bR and flavin were deposited onto the patterned (9-pixelized) ITO glass by LB technique. A 9-pixel biomolecular photoreceptor was fabricated with a sandwich-type structure of ITO/LB films/electrolyte gel/Pt. Since each functional molecule shows its own response characteristic according to the light illumination in the visible region, the simplified knowledge-based algorithm for interpretation of the incident light wavelength (color) was proposed based on the basic rule describing the relationship between the photoelectric response characteristics and the incident light wavelength. When simple color images were projected onto the photoreceptor, the primary colors in visible light region, red, green, and blue were clearly recognized, and the projected color images were fairly well reproduced onto the color monitor by the proposed photoreceptor with the knowledge-based algorithm. It is concluded that the proposed device has a capability of recognizing the color images and can be used as a model system to simulate the information processing function of the human visual system.     

Improved Isolation Procedures for the Purple Membrane of Halobacterium Halobium

Techniques for purifying the purple membrane of Halobacterium halobium are given. This purple membrane contains a chromoprotein with a retinal prosthetic group similar to rhodopsin, the chromoprotein found in the visual systems of higher invertebrates and vertebrates. The described purple membrane isolation procedures yield a highly purified preparation as determined by transmitting electron microscopy and gel electrophoresis. Critical analysis of the absorption spectra of the purple membrane was also employed to establish criteria of purity for the preparation. The visible absorption spectra of the purified purple membrane preparation in buffer was found to have a maximum at 559 nm which shifted to 567 nm on light exposure. No indication of any spectral perturbation arising from bacterioruberin-containing membrane, the major contaminant in purple membrane preparations, was found. Furthermore, the ratio of protein aromatic amino acid absorbance at 280 nm to chromophore absorbance at 567 nm was found to be 1.5 in light-exposed preparations compared to the previously reported ratio of 2.0.³ The decrease in the value of this ratio is also indicative of an increase in the purity of the purple membrane preparation.     

Purinergic signalling in the subretinal space: a role in the communication between the retina and the RPE

The retinal pigment epithelium (RPE) is separated from the photoreceptor outer segments by the subretinal space. While the actual volume of this space is minimal, the communication that occurs across this microenvironment is important to the visual process, and accumulating evidence suggests the purines ATP and adenosine contribute to this communication. P1 and P2 receptors are localized to membranes on both the photoreceptor outer segments and on the apical membrane of the RPE which border subretinal space. ATP is released across the apical membrane of the RPE into this space in response to various triggers including glutamate and chemical ischemia. This ATP is dephosphorylated into adenosine by a series of ectoenzymes on the RPE apical membrane. Regulation of release and ectoenzyme activity in response to light-sensitive signals can alter the balance of purines in subretinal space, and thus coordinate communication across subretinal space with the visual process.