Improved Success of Sparse Matrix Protein Crystallization Screening with Heterogeneous Nucleating Agents

被引:51
作者
Thakur, Anil S. [1 ]
Robin, Gautier [2 ,3 ]
Guncar, Gregor [1 ]
Saunders, Neil F. W. [1 ]
Newman, Janet [4 ]
Martin, Jennifer L. [1 ,2 ,3 ]
Kobe, Bostjan [1 ,2 ,3 ]
机构
[1] Univ Queensland, Sch Mol & Microbial Sci, Brisbane, Qld, Australia
[2] Univ Queensland, Inst Mol Biosci, Brisbane, Qld, Australia
[3] Univ Queensland, Australian Res Council ARC, Special Res Ctr Funct & Appl Genom, Brisbane, Qld, Australia
[4] Commonwealth Sci & Ind Res Org, Mol & Hlth Technol, Parkville, Vic, Australia
来源
PLOS ONE | 2007年 / 2卷 / 10期
基金
澳大利亚研究理事会;
关键词
D O I
10.1371/journal.pone.0001091
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background. Crystallization is a major bottleneck in the process of macromolecular structure determination by X-ray crystallography. Successful crystallization requires the formation of nuclei and their subsequent growth to crystals of suitable size. Crystal growth generally occurs spontaneously in a supersaturated solution as a result of homogenous nucleation. However, in a typical sparse matrix screening experiment, precipitant and protein concentration are not sampled extensively, and supersaturation conditions suitable for nucleation are often missed. Methodology/Principal Findings. We tested the effect of nine potential heterogenous nucleating agents on crystallization of ten test proteins in a sparse matrix screen. Several nucleating agents induced crystal formation under conditions where no crystallization occurred in the absence of the nucleating agent. Four nucleating agents: dried seaweed; horse hair; cellulose and hydroxyapatite, had a considerable overall positive effect on crystallization success. This effect was further enhanced when these nucleating agents were used in combination with each other. Conclusions/Significance. Our results suggest that the addition of heterogeneous nucleating agents increases the chances of crystal formation when using sparse matrix screens.
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页数:6
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