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首页> 外文期刊>Acta crystallographica. Section D, Structural biology >The copper(II)-binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography
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The copper(II)-binding tripeptide GHK, a valuable crystallization and phasing tag for macromolecular crystallography

机译：铜(II)绑定三肽原则为,有价值结晶和逐步标记大分子晶体学

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摘要

The growth of diffraction-quality crystals and experimental phasing remain two of the main bottlenecks in protein crystallography. Here, the high-affinity copper(II)-binding tripeptide GHK was fused to the N-terminus of a GFP variant and an MBP-FG peptide fusion. The GHK tag promoted crystallization, with various residues (His, Asp, His/Pro) from symmetry molecules completing the copper(II) square-pyramidal coordination sphere. Rapid structure determination by copper SAD phasing could be achieved, even at a very low Bijvoet ratio or after significant radiation damage. When collecting highly redundant data at a wavelength close to the copper absorption edge, residual S-atom positions could also be located in log-likelihood-gradient maps and used to improve the phases. The GHK copper SAD method provides a convenient way of both crystallizing and phasing macromolecular structures, and will complement the current trend towards native sulfur SAD and MR-SAD phasing.
机译：diffraction-quality晶体的生长实验逐步保持两个主要的瓶颈在蛋白质晶体学。高亲和性铜(II)绑定三肽原则为融合GFP变体的n端和一个MBP-FG肽融合。结晶,各种残留物(他的,Asp的,他/ Pro)对称分子完成铜(II) square-pyramidal协调球体。快速结构测定铜悲伤逐步实现,即使在非常低的水平毕吉伯比率或之后重要的辐射损害。波长接近铜吸收边缘,剩余S-atom位置也可以在log-likelihood-gradient地图和习惯提高阶段。提供了一个方便的方法的结晶,并将逐步大分子结构补充当前本地的趋势硫悲伤和MR-SAD定相。

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