温度对脂质囊泡中单体菌紫质分子结构和光电响应的影响

本实验用人工双分子平板膜系统(BLM)测量了紫膜碎片和在DMPC脂质襄泡膜中的单体菌紫质分子的光电响应以及与温度的关系(处理温度17℃至31℃).温度对紫膜碎片的光电响应影响不大,但对单体菌紫质分子的光电响应有明显影响.用园二色(CD)方法相应地观察了温度对紫膜碎片和单体菌紫质分子在可见波长范围内的CD谱的影响 同样观察到温度对单体菌紫质分子的CD谱有明显影响.两者的影响很可能与脂质襄泡中DMPC的相变温度有关.     

缬氨霉素以及膜电位对菌紫质分子在脂质囊泡膜中运动的影响

用瞬间二向色性方法测量了菌紫质分子在DMPC脂质囊泡膜中的旋转扩散运动.观察了缬氨霉素和膜电位对菌紫质分子旋转扩散运动的影响.在低脂与蛋白比例时,缬氨霉素明显影响菌紫质分子的旋转扩散运动,在高脂与蛋白比例时,缬氨霉素对菌紫质分子的旋转扩散动的影响不明显.无论在高的或低的脂与蛋白比例下,膜电位都影响到菌紫质分子的旋转扩散运动.     

菌紫质分子的重组对结构及运动的影响

本实验用闪光诱导的瞬间二向色性方法分别测量了用全反视黄醛重组以及用四溴荧光素标记的菌紫质分子在脂质囊泡膜中的旋转扩散运动以及吸收过程随时间的变化(寿命).用全反视黄醛重组后不影响菌紫质分子在膜中的旋转扩散运动,它们与蜂毒的相互作用也无明显差别.用四溴荧光素标记的菌紫质分子的寿命很短,不可能用于测量菌紫质分子在膜中的旋转运动.     

外加电场对平板膜(BLM)中单体菌紫质分子bR光电响应的影响

本文研究了外加电场对单体菌紫质分子在平板膜系统中光电响应的影响.外加正电场时(外池为正,内池为负),光电响应信号变小,电场加至+52mV时,光电响应信号极性发生反转.外加负电场时,光电响应信号增大.本文提出了极性发生反转的可能模型,并对其机理进行了初步探讨.     

切割菌紫质分子C端多肽对脂质泡中菌紫质分子在膜中旋转运动的影响

本实验用闪光诱导的瞬间二向色性方法分别测量了重组在DMPC脂质泡中的菌紫质分子和用木瓜蛋白酶切割C端多肽的菌紫质分子的旋转扩散运动。酶切的菌紫质分子与原菌紫质分子相比 前者旋转扩散运动加快,不可动分子的比例下降。在蜂毒或乙酰蜂毒存在下,酶切的菌紫质分子旋转扩散运动也受到较少的阻碍     

温度及膜中蛋白质含量与膜粘度和膜中分子排列方向的关系

本实验用闪光诱导的瞬间二向色性方法测量了不同温度以及不同蛋白质含量下菌紫质分子在脂质囊泡膜中的旋转扩散运动.根据旋转扩散运动得到了温度和蛋白质含量与膜粘度以及分子在膜中排列方向的关系.温度和蛋白质的含量都影响膜的粘度,但并不影响蛋白质分子在膜中的排列方向.     

温度及膜中蛋白质含量与膜粘度和膜中分子排列方向的关系

本实验用闪光诱导的瞬间二向色性方法测量了不同温度以及不同蛋白质含量下菌紫质分子在脂质囊泡膜中的旋转扩散运动.根据旋转扩散运动得到了温度和蛋白质含量与膜粘度以及分子在膜中排列方向的关系.温度和蛋白质的含量都影响膜的粘度,但并不影响蛋白质分子在膜中的排列方向.     

菌紫质LB膜时空信号响应模型及方向检测效应的研究

在测量和分析了菌紫质ＬＢ膜的时空信号响应的基础上,提出了菌紫质ＬＢ膜的时空信号响应模型。并用菌紫质ＬＢ膜的时空信号响应模型。并用菌紫质ＬＢ膜的这些特性设计了能检测“对比边”运动方向、角度和转速的方法。     

菌紫质LB膜时空信号响应模型及方向检测效应的研究

在测量和分析了菌紫质ＬＢ膜的时空信号响应的基础上,提出了菌紫质ＬＢ膜的时空信号响应模型。并用菌紫质ＬＢ膜的这些特性设计了能检测“对比边”运动方向、角度和转速的方法。     

紫膜Langmuir-Blodgett膜的光电压

八十年代生物分子电子学和生物计算机研究的兴起,嗜盐菌紫膜蛋白菌紫质由于其结构稳定,当受到光照时,能发生同素异构变化,具有光色互变和光驱动质子泵功能、双稳态特性以及皮秒到毫秒级的光电响应特性,使其成为目前国内外关注的研究生物分子电子器件的理想材料之一,应用前途是发展生物分子器件和生物芯片等。由于紫膜具有不对称性而菌紫质的光驱动质子泵具有方向性,因而必须有序组装而且可以有序组装。紫膜     

pH梯度对平板膜中单体菌紫质分子光电响应的影响

利用重组技术把单体bR包入DMPC脂质囊泡中,通过吸收光谱和圆二色谱的测定,证明bR在囊泡中仍以单体存在。同时测定了外界pH梯度对单体bR脂质囊泡BLM光电响应的影响,观察到pH值当内池大于外池其差值超过2时就可产生光电响应信号极性的反转,并对此结果作了一定的讨论。     

脂质泡中的菌紫质旋转运动的介质依赖性

用闪光诱导瞬间二向色性方法测量了蔗糖和甘油对菌紫质（ＢＲ）分子在脂质泡中的旋转扩散运动的影响。结果表明脂质囊泡本身在悬浮液中的运动并不会影响ＢＲ分子在膜中的旋转运动的测量。蔗糖和甘油对ＢＲ旋转运动的影响在相变温度以上和相变温度以下是不同的,相变温度以下的主要作用是相分离,使运动减慢,相变温度以上的作用可能是脂的分散,使运动加快。     

Electric response of a back photoreaction in the bacteriorhodopsin photocycle.

The electric response of a back photoreaction in the bacteriorhodopsin photocycle was investigated. The proton pumping activity of green flash excited bacteriorhodopsin stops if the M412 form is illuminated by blue light (Karvaly and Dancsházy, 1977). In the present work a fast negative displacement current signal was measured in an oriented membrane suspension system, indicative of back movement of protons from M412 to BR570. Quantitative evaluation of the data shows that there are at least two steps in the back reaction, with different rate constants. The temperature dependence of the rate constants show simple linear Arrhenius behavior between 5 degree and 40 degree C. The rate constants were slower by a factor of 1.8 in D2O suspension. The relevance of the protein electric response signals (PERS) observed in this paper to the early receptor potential is discussed.     

Flash kinetic study of the last steps in the photoinduced reaction cycle of bacteriorhodopsin

The reaction cycle of light adapted bacteriorhodopsin (BR) in aqueous purple membrane suspensions was studied by laser flash photolysis at different temperatures (2–49°C) and pH values (3–10). The activation energy for several reaction steps was determined at pH 7.6. The kinetics of O-bacteriorhodopsin (one of the last intermediates in the cycle) were analyzed in some detail and it was found that the simple consecutive reaction scheme M-BR → O-BR → BR may explain the kinetics of O-bacteriorhodopsin as measured at 680 nm. Since the pH change in neutral aqueous suspensions of purple membrane follows a similar kinetics as O-bacteriorhodopsin it is suggested that protons are released during the reaction M-BR → O-BR and taken up again during the reaction O-BR → BR.Another long-lived intermediate, which absorbs to a greater extent than bacteriorhodopsin at 570 nm and less than bacteriorhodopsin at 420 nm, was identified with the strongly fluorescing species, pseudo- or P-bacteriorhodopsin. The decay of P-bacteriorhodopsin in bacteriorhodopsin had an activation energy of only approx. 1.2 kcal/mol, which suggests that the last step of the photocycle is a relaxation around a single bond.At pH 9–10, the simple first-order kinetics of all the intermediates were changed into a kinetics consisting of two first-order decays. This change of kinetics was accompanied by a drastic decrease in the rotational diffusion relaxation time.To explain the results obtained in this work and those of others, a model involving proton uptake and release by the Schiff base nitrogen combined with an isomerization reaction is finally proposed.