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Protein Self-Organization In Vitro and In Vivo: Partitioning between Physical Biochemistry and Cell Biology

机译:体内和体外蛋白质自组织:物理生化与细胞生物学之间的划分

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摘要

Protein folding is a hierarchical process, driven by the accumulation of increments of free energy from local interactions between neighboring residues, secondary structural elements, domains and subunits. The latter represent independent folding units. Thus, the folding kinetics divide into the collapse of sub-domains and domains and their merging to form the compact tertiary fold. In proceeding to oligomeric proteins, docking of structured monomers is the final step. In agreement with this mechanism, in vitro experiments show that the overall mechanism of folding and association obeys uni-bimolecular kinetics with aggregation as a competing side reaction. In vivo, accessory proteins serve to shift the kinetic partitioning between assembly and misassembly toward the native state. So far, co- and post-translational protein folding in the cell has been withstanding a detailed kinetic analysis. Despite obvious differences between the crowded cytosol and optimized in vitro folding conditions, the general mechanism of protein self-or-ganization within and without the cell seems to be similar. Effects of solvent parameters on the rate and mode of protein folding are less significant than predicted. Addition of small ligands and compatible solutes allow nucleation steps and viscosity effects to be analyzed. The absence of chimeras after synchronous in vitro reconstitution of oligomeric enzymes proves subunit interactions to be highly specific.
机译:蛋白质折叠是一个分层的过程,由相邻残基,二级结构元件,结构域和亚基之间的局部相互作用所产生的自由能增量的累积驱动。后者代表独立的折叠单元。因此,折叠动力学分为亚结构域和结构域的折叠以及它们的合并以形成紧凑的第三折叠。在进行寡聚蛋白的过程中,结构化单体的对接是最后一步。与这种机制一致,体外实验表明,折叠和缔合的整体机制遵循单双分子动力学,而聚集是竞争性副反应。在体内,辅助蛋白用于将组装和错误组装之间的动力学分配移向天然状态。到目前为止,细胞中的共翻译和翻译后蛋白质折叠已经经受了详细的动力学分析。尽管拥挤的细胞质与优化的体外折叠条件之间存在明显差异,但在细胞内外的蛋白质自组织的一般机制似乎是相似的。溶剂参数对蛋白质折叠速率和模式的影响不如预期的重要。添加小的配体和相容的溶质可以分析成核步骤和粘度效应。同步体外重组寡聚酶后不存在嵌合体,证明亚基相互作用具有高度特异性。

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