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Self-Assembly of Protein and Liquid-Liquid Phase Separation under Shear Stress : Investigations by Phase Contrast Rheoscope

剪切应力下蛋白质的自组装和液-液相分离：通过相差流变仪进行研究

基本信息

批准号：
07640774
负责人：
KAIBARA Kozue
金额：
$ 1.47万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

1.Liquid-Liquid Phase Separation in Elastomeric Protein-Water System : Examinations of Critical CharacteristicsThe most remarkable characteristics of elastomeric protein-water system, temperature-dependent coacervation, can be treated as a liquid-liquid phase separation process with the lower critical solution temperature. For the bovine neck ligamental alpha-elastin-water system, near the concentration of 0.11 mg/ml and the temperature of 21.5ﾟC,critical divergences of the forward scattering intensity and the correlation length were observed by dynamic light scattering experiments. In the present study, critical and off-critical characteristics of the phase separation of alpha-elastin-water system were examined by the light scattering and viscometric measurements as well as by the phase contrast microscopic observations. The phase contrast microscopic observations were useful for the system with high concentration of alpha-elastin and for the late stages of phase separation process.2. … More Assemblies of Phase Contrast RheoscopeSelf-assembly process of elastin precursor, tropoelastin, can be mimicked by the physicochemical molecular aggregation process during the temperature-dependent coacervation of elastomeric protein water system. However, the biological process proceeds in the extracellular space where extremely unusual conditions are realized under shear stress and structural disturbance of solvent. To examine the phase behavior in these conditions, rheoscope was introduced by assembling inverted phase contrast microscope and cone-plate rotary viscometer. Basic arrangements of the rheoscope is that the viscometer with transparent cone attachment is placed on the stage with horizontally positioning of the light sauce and condenser. The rheoscope is capable to measure viscosity and simultaneously observe phase behavior by monitoring the appearances of microcoacervate droplets and by tracing their size and shape transformations. The most noticeable advantages of rheoscope is observations of the phase separation process under regulated shear stress as a model space of the extracellular crevice. Less

1.弹性蛋白质-水体系的液-液相分离：临界特性的考察弹性蛋白质-水体系最显著的特性是温度依赖性凝聚，它可以看作是具有较低临界溶解温度的液-液相分离过程。对于牛颈韧带α-弹性蛋白-水体系，在浓度为0.11mg/ml，温度为21.5 ℃附近，动态光散射实验观察到前向散射强度和相关长度的临界发散。在本研究中，α-弹性蛋白-水系统的相分离的临界和非临界特性进行了研究，通过光散射和粘度测量以及相差显微镜观察。相差显微镜观察对高浓度α-弹性蛋白体系和相分离过程的后期阶段是有用的. ...更多信息相衬流变仪的组装弹性蛋白前体原弹性蛋白的自组装过程可以通过弹性蛋白-水体系的温度依赖性凝聚过程中的物理化学分子聚集过程来模拟。然而，生物过程在细胞外空间中进行，其中在剪切应力和溶剂的结构干扰下实现极不寻常的条件。采用倒置相衬显微镜和锥板旋转粘度计组合的流变仪考察了该条件下的相行为。该流变仪的基本布置是将带有透明锥附件的粘度计放置在载物台上，光酱料和聚光镜水平放置。流变仪能够测量粘度，同时通过监测微凝聚层液滴的外观并通过跟踪它们的尺寸和形状转变来观察相行为。流变仪最显著的优点是作为细胞外裂隙的模型空间，可以观察在调节的剪切应力下的相分离过程。少