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利用吸附和解吸同时提高蛋白质结晶成功率和晶体质量。

Utilisation of adsorption and desorption for simultaneously improving protein crystallisation success rate and crystal quality.

作者信息

Guo Yun-Zhu, Sun Li-Hua, Oberthuer Dominik, Zhang Chen-Yan, Shi Jian-Yu, Di Jiang-Lei, Zhang Bao-Liang, Cao Hui-Ling, Liu Yong-Ming, Li Jian, Wang Qian, Huang Huan-Huan, Liu Jun, Schulz Jan-Mirco, Zhang Qiu-Yu, Zhao Jian-Lin, Betzel Christian, He Jian-Hua, Yin Da-Chuan

机构信息

1] Key Laboratory for Space Bioscience &Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an 710072, Shaanxi, PR China [2] Chang'an University, Middle Section of Nan Erhuan Road, Xi'an City, Shaanxi, PR China.

Shanghai Institute of Applied Physics, Chinese Academy of Sciences, 239 Zhangheng Road, Shanghai 201204, P.R. China.

出版信息

Sci Rep. 2014 Dec 4;4:7308. doi: 10.1038/srep07308.

Abstract

High-quality protein crystals of suitable size are an important prerequisite for applying X-ray crystallography to determine the 3-dimensional structure of proteins. However, it is often difficult to obtain protein crystals of appropriate size and quality because nucleation and growth processes can be unsuccessful. Here, we show that by adsorbing proteins onto porous polystyrene-divinylbenzene microspheres (SDB) floating on the surface of the crystallisation solution, a localised high supersaturation region at the surface of the microspheres and a low supersaturation region below the microspheres can coexist in a single solution. The crystals will easily nucleate in the region of high supersaturation, but when they grow to a certain size, they will sediment to the region of low supersaturation and continue to grow. In this way, the probability of crystallisation and crystal quality can be simultaneously increased in a single solution without changing other crystallisation parameters.

摘要

高质量且尺寸合适的蛋白质晶体是应用X射线晶体学来确定蛋白质三维结构的重要前提。然而,由于成核和生长过程可能不成功,通常很难获得尺寸和质量合适的蛋白质晶体。在这里,我们表明,通过将蛋白质吸附到漂浮在结晶溶液表面的多孔聚苯乙烯 - 二乙烯基苯微球(SDB)上,微球表面的局部高过饱和区域和微球下方的低过饱和区域可以在单一溶液中共存。晶体将很容易在高过饱和区域成核,但当它们生长到一定大小时,会沉淀到低过饱和区域并继续生长。通过这种方式,在不改变其他结晶参数的情况下,可以在单一溶液中同时提高结晶概率和晶体质量。

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