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通过电子自旋共振(ESR)测量共价结合自旋标记的大分子旋转相关时间

[Macromolecule rotative correlation time measurement by ESR for covalently bound spin label].

作者信息

Dudich I V, Timofeev V P, Vol'kenshteĭn M V, Misharin A Iu

出版信息

Mol Biol (Mosk). 1977 May-Jun;11(3):685-93.

PMID:223033
Abstract

The dependence from temperature and viscosity of the shifts of the internal and external wide extremums in the ESR spectra of spin labelled bovine serum albumin has been studied. 2,2,6,6-tetramethylpiperidine-NI-oxyl-4-iodacetamide was used as a spin label. The obtained dependences was shown to be a consequence of the label participation in two types of rotations: an anisotropic fast rotation with tau less than 10(-9) sec relatively to a macromolecule, and the isotropic one with tau greater than 10(-8) sec due to rotation of the macromolecule itself. These conclusions were done on the basis of a model for complex rotation of the spin label. Comparison of theoretical and experimental data makes it possible to determined the correlation time for the protein moiety, to evaluate quantitatively the polarity of surroundings of the iminoxyl and to introduce a numerical parameter for the degree of mobility of the spin label relatively to protein molecule.

摘要

研究了自旋标记牛血清白蛋白的电子自旋共振(ESR)谱中内部和外部宽极值位移对温度和粘度的依赖性。使用2,2,6,6-四甲基哌啶-NI-氧基-4-碘乙酰胺作为自旋标记。结果表明,所获得的依赖性是标记参与两种旋转类型的结果:相对于大分子,τ小于10^(-9)秒的各向异性快速旋转,以及由于大分子自身旋转导致的τ大于10^(-8)秒的各向同性旋转。这些结论是基于自旋标记的复杂旋转模型得出的。理论数据与实验数据的比较使得确定蛋白质部分的相关时间、定量评估亚胺氧基周围环境的极性以及引入自旋标记相对于蛋白质分子的迁移程度的数值参数成为可能。

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