Utilization of Cyclodextrins and Its Derivative Particles as Nucleants for Protein Crystallization,Crystal Growth & Design

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Utilization of Cyclodextrins and Its Derivative Particles as Nucleants for Protein Crystallization
Crystal Growth & Design ( IF 3.2 ) Pub Date : 2017-10-31 00:00:00 , DOI: 10.1021/acs.cgd.7b00455
Xue-Zhou Yang ₁ , Chen-Yan Zhang ₁ , Qian-Jin Wang ₂ , Yun-Zhu Guo ₁ , Chen Dong ₁ , Er-Kai Yan ₁ , Wen-Jing Liu ₁ , Xi-Wang Zheng ₁ , Da-Chuan Yin ₁

Affiliation

Finding new nucleants to promote protein crystallization is an important task, especially for purposes other than structural determination. Here, we investigated cyclodextrins and its derivative particles, as potential nucleants for protein crystallization. β-Cyclodextrin (β-CD) and its derivatives (including p-toluenesulfonyl-β-cyclodextrin (PTCD), polymer-β-cyclodextrin (PCD), mono-(6-(1,6-hexamethylenediamine)-6-deoxy)-β-cyclodextrin (MHCD) and mercapto-β-cyclodextrin (MCD)) were used as nucleants. The experimental results confirmed that β-CD and its derivatives showed significantly positive effects, promoting protein crystallization and improving crystal quality. A larger number of protein molecules (including lysozyme, catalase, subtilisin A VIII, concanavalin A VI, α-chymotrypsinogen, proteinase K, cellulase, papain, glucose isomerase, hemoglobin, and ribonuclease A XII) attached to the particles usually corresponded to a higher crystallization success rate. More detailed analysis showed that cyclodextrins exhibited the best performance when the overall charge of protein in solution was the opposite to zeta potential of the cyclodextrins particle. Our results showed that cyclodextrins can be useful as nucleants due to the ease of modifying them to suit the crystallization of different proteins, and they can be explored for use in the mass purification of proteins for the biopharmaceutical industry. Furthermore, the phenomenon discovered in this study pointed toward a way to find new nucleants based on the overall charge of proteins in a solution: the nucleants should preferably be the opposite between the overall charge of target protein and the zeta potential of the cyclodextrin particle.

中文翻译：

环糊精及其衍生物作为核酸结晶的成核剂

寻找新的成核剂以促进蛋白质结晶是一项重要的任务，特别是出于结构确定以外的目的。在这里，我们研究了环糊精及其衍生物颗粒，作为蛋白质结晶的潜在成核剂。β-环糊精（β-CD）及其衍生物（包括p-甲苯磺酰基-β-环糊精（PTCD），聚合物-β-环糊精（PCD），单-（6-（1,6-六亚甲基二胺）-6-脱氧）-β-环糊精（MHCD）和巯基-β-环糊精（ MCD））用作成核剂。实验结果证实，β-CD及其衍生物显示出明显的积极作用，促进了蛋白质结晶并改善了晶体质量。附着在颗粒上的蛋白质分子较多（包括溶菌酶，过氧化氢酶，枯草杆菌蛋白酶A VIII，伴刀豆球蛋白A VI，α-胰凝乳蛋白酶原，蛋白酶K，纤维素酶，木瓜蛋白酶，葡萄糖异构酶，血红蛋白和核糖核酸酶A XII）通常对应于较高的结晶成功率。更详细的分析表明，当溶液中蛋白质的总电荷与环糊精颗粒的Zeta电位相反时，环糊精表现出最佳性能。我们的结果表明，环糊精由于易于修饰以适合不同蛋白质的结晶而可用作成核剂，并且可以探索它们用于生物制药行业蛋白质的大规模纯化。此外，在这项研究中发现的现象指出了一种基于溶液中蛋白质的总电荷寻找新的成核剂的方法：成核剂应优选在目标蛋白质的总电荷与环糊精颗粒的ζ电势之间相反。

更新日期：2017-11-01

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