Adsorption of protein onto dye-ligand affinity adsorbent: Molecular dynamics simulation and experimental study

蛋白质在染料-配体亲和吸附剂上的吸附：分子动力学模拟和实验研究

• 批准号: 245147205
• 负责人: Professor Dr.-Ing. Georg Fieg
• 金额: --
• 依托单位: Institut für Systemverfahrenstechnik V-4
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/245147205?language=en
• 关键词: Adsorption; protein; onto; dye; ligand

Affinity chromatography is a well-established method for the identification, purification, and separation of macromolecules based on highly specific molecular recognition. Dye ligand affinity chromatography has been widely utilized for protein purification. Immobilized textile triazine dyes, particularly Cibacron blue 3GA (CB) has been used as affinity chromatography tools for protein purification for a long time. CB is able to bind various proteins. However, its interaction with a large number of seemingly unrelated proteins inevitably compromises its protein binding specificity and endows these molecules with a serious drawback. For an improved application of textile dyes in the separation of proteins, it is very important to understand the binding and specific mechanisms between dye ligand and proteins. In this project, serum albumin and hemoglobin which have been investigated in our previous works will be further studied as model proteins to investigate their adsorption onto dye ligand (CB) affinity chromatographic media. Due to the lack of experimental method that can detect the dynamic adsorption process within adsorbent pores, molecular dynamics (MD) simulations with both coarse grained and all atom models will be used for the first time in this study to investigate the dye ligand affinity chromatographic adsorption of proteins on a molecular scale. On the basis of the results of MD simulations and the former experimental research, both single and binary adsorption experiments of these proteins onto dye ligand affinity adsorbent will be performed under conditions which are rationally designed according to the binding mechanism learned from the MD simulations. A combination of MD simulations and adsorption experiments will give a comprehensive understanding of the binding and specific mechanisms of the affinity adsorption from both microscopic and macroscopic point of view. This understanding is a profound basis for the design and screening of new affinity dye ligands, as well as the design, optimization and control of the dye ligand affinity chromatography of proteins.

亲和层析是一种基于高度特异的分子识别来鉴定、纯化和分离大分子的成熟方法。染料配基亲和层析已被广泛应用于蛋白质的纯化。长期以来，固定化纺织三嗪染料，特别是Cibacron Blue 3GA(CB)一直被用作亲和层析工具来纯化蛋白质。CB能与多种蛋白质结合。然而，它与大量看似无关的蛋白质的相互作用不可避免地损害了它与蛋白质结合的特异性，并赋予这些分子一个严重的缺陷。为了提高纺织染料在蛋白质分离中的应用，了解染料配体与蛋白质之间的结合及其特定的机理是非常重要的。在本项目中，我们将进一步研究我们以前工作中研究过的血清白蛋白和血红蛋白作为模型蛋白，以考察它们在染料配体(CB)亲和层析介质上的吸附情况。由于目前尚无实验方法来研究吸附树脂孔内的动态吸附过程，本研究首次采用粗粒模型和全原子模型的分子动力学(MD)模拟方法，从分子水平上研究了染料配基亲和层析对蛋白质的吸附作用。在分子动力学模拟结果和前人实验研究的基础上，根据分子动力学模拟得到的结合机理，在合理设计的条件下，分别进行了染料配基亲和吸附剂对这些蛋白质的单吸附和二元吸附实验。分子动力学模拟和吸附实验相结合，将从微观和宏观两个角度对亲和吸附的结合和具体机理有一个全面的了解。这一认识为新型亲和染料配基的设计和筛选，以及蛋白质亲和色谱染料配基的设计、优化和控制奠定了基础。