Liquid-liquid phase separation and crystallization in protein solutions induced by multivalent metal ions

多价金属离子诱导蛋白质溶液中的液-液相分离和结晶

• 批准号: 197073534
• 负责人: Privatdozent Dr. Fajun Zhang, Ph.D.
• 金额: --
• 依托单位: Lehrstuhl Physik der molekularen und biologischen Materie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/197073534?language=en
• 关键词: Liquid; liquid; phase; separation; crystallization

Liquid-liquid phase separation (LLPS) in protein solutions (i.e. the separation into two liquid phases with low and high protein concentration, respectively) is a crucial phenomenon for protein crystallization as well as protein-association related diseases, such as sickle cell anemia and cataracts. However, the physical mechanisms on the condition of LLPS and its effect on subsequent protein crystallization are still poorly understood. In this proposal, we will undertake a comprehensive study of protein solutions in the presence of multivalent metal ions, where a reentrant condensation phase behavior and LLPS have been observed recently in our group. Our aim is to explore the fundamental mechanisms as well as the key parameters behind of this phenomenon on a molecular level and finally to control the phase behavior of protein solutions. The following questions will be addressed: (a) What is the effective protein-protein interaction potential when LLPS occurs? (b) What are the key parameters determining the phase behavior of protein solutions (nature and concentration of salt, pH, T, …)? (c) What are the differences and similarities between LLPS induced by multivalent ions vs polymer (PEG)? A set of complementary experimental techniques including small-angle scattering (SAXS, SANS), static/dynamic light scattering (SLS/DLS), and spectroscopy (FTIR, UV-vis and circular dichroism) will be used to address the different issues in these complex systems.

蛋白质溶液中的液-液相分离（即分离成蛋白浓度分别为低和高的两种液相）是蛋白质结晶以及镰状细胞性贫血、白内障等蛋白质相关疾病的重要现象。然而，LLPS条件下的物理机制及其对后续蛋白质结晶的影响仍然知之甚少。在本提案中，我们将对多价金属离子存在下的蛋白质溶液进行全面研究，其中我们小组最近观察到可重入缩合相行为和LLPS。我们的目标是在分子水平上探索这种现象背后的基本机制和关键参数，并最终控制蛋白质溶液的相行为。将解决以下问题：(a) LLPS发生时，有效的蛋白质-蛋白质相互作用潜力是什么？(b)决定蛋白质溶液相行为的关键参数是什么（盐的性质和浓度、pH、T等）？(c)多价离子诱导的LLPS与聚合物（PEG）诱导的LLPS有何异同？一套互补的实验技术，包括小角度散射（SAXS， SANS），静态/动态光散射（SLS/DLS）和光谱学（FTIR， UV-vis和圆二色）将被用来解决这些复杂系统中的不同问题。

项目成果

Ion-activated attractive patches as a mechanism for controlled protein interactions

离子激活的吸引斑块作为受控蛋白质相互作用的机制

• DOI: 10.1038/srep07016
• 发表时间: 2014
• 期刊: Scientific Reports
• 影响因子: 4.6
• 作者: F. Roosen-Runge;F. Zhang;F. Schreiber;R. Roth
• 通讯作者: R. Roth

Charge-controlled metastable liquid-liquid phase separation in protein solutions as a universal pathway towards crystallization

蛋白质溶液中电荷控制的亚稳态液-液相分离作为结晶的通用途径

• DOI: 10.1039/c2sm07008a
• 发表时间: 2012
• 期刊: Soft Matter
• 影响因子: 3.4
• 作者: F. Zhang;R. Roth;M. Wolf;F. Roosen-Runge;M. W. A. Skoda;R. M. J. Jacobs;R. Roth;M. Sztucki;F. Schreiber
• 通讯作者: F. Schreiber

Nonclassical Pathways of Protein Crystallization in the Presence of Multivalent Metal Ions

• DOI: 10.1021/cg501099d
• 发表时间: 2014-12-01
• 期刊: CRYSTAL GROWTH & DESIGN
• 影响因子: 3.8
• 作者: Sauter, Andrea;Oelker, Melanie;Schreiber, Frank
• 通讯作者: Schreiber, Frank

Effective interactions in liquid-liquid phase separated protein solutions induced by multivalent ions

多价离子诱导的液-液相分离蛋白质溶液中的有效相互作用

• DOI: 10.15496/publikation-4822
• 发表时间: 2014
• 作者: Wolf M.

Protein cluster formation in aqueous solution in the presence of multivalent metal ions--a light scattering study.

• DOI: 10.1039/c3sm52447g
• 发表时间: 2014-01
• 期刊: Soft matter
• 影响因子: 3.4
• 作者: Daniel Soraruf;F. Roosen‐Runge;M. Grimaldo;F. Zanini;R. Schweins;T. Seydel;Fajun Zhang;R. Roth;M. Oettel;F. Schreiber