Ordered self-assembly of polypeptide fragments to form nativelike dimeric trp repressor.
Ordered self-assembly of polypeptide fragments to form nativelike dimeric trp repressor.
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DOI：
10.1126/science.1736361
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发表时间：
1992-01
期刊：
影响因子：
56.9
通讯作者：
Maria Luisa Tasayco;Jannette Carey
中科院分区：
文献类型：
--
作者：
Maria Luisa Tasayco;Jannette Carey
已经产生了大肠杆菌 trp 阻遏物的亚结构域大小的蛋白水解片段，这些片段按照确定的顺序组装以再生完全天然的二聚体。通过表征分离和重组片段的二级和三级结构，组装中间体的结构可以与从折叠速率的光谱测量推导出的完整阻遏物的动力学折叠途径相关联。这些中间体的天然结构进一步证明蛋白质折叠途径反映了天然状态下发现的二级结构单元和组装体的稳定性。蛋白水解方法通常可用于向光谱确定的折叠机制添加结构细节。
Subdomain-size proteolytic fragments of Escherichia coli trp repressor have been produced that assemble in defined order to regenerate fully native dimers. By characterization of the secondary and tertiary structures of isolated and recombined fragments, the structure of assembly intermediates can be correlated with the kinetic folding pathway of the intact repressor deduced from spectroscopic measurement of folding rates. The nativelike structure of these intermediates provides further evidence that protein folding pathways reflect the stabilities of secondary structural units and assemblies found in the native state. The proteolytic method should be generally useful in adding structural detail to spectroscopically determined folding mechanisms.