Mechanism of Formation and Functional Properties of Lipoprotein Discs Stabilized by Amphiphilic Maleic Acid-Containing Alternating Copolymers

两亲性马来酸交替共聚物稳定脂蛋白盘的形成机制和功能特性

• 批准号: 388943162
• 负责人: Professor Dr. Heinz-Jürgen Steinhoff
• 金额: --
• 依托单位: Russian Foundation for Basic Research
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/388943162?language=en
• 关键词: Mechanism; Formation; Functional; Properties; Lipoprotein

Amphiphilic maleic acid-containing alternating copolymers (MACPs), in which units of maleic acid alternate with (variable) hydrocarbon units, account for a recent major methodical breakthrough in the study of membrane proteins. Several MACPs were found to solubilize membrane proteins both from artificial and natural lipid bilayers yielding discoidal MACP-encased lipid/lipoprotein particles with uniform diameter in the range from 10-30 nm. Enzymes seem to remain functional within MACP-particles; therefore, such particles make even those membrane proteins, which are unstable in the presence of detergent, accessible to techniques that require homogenous water-soluble (single) particles. It remains, however, unclear how MACPs extract membrane proteins in the absence of detergents, how the different components of the MACP-lipid/lipoprotein particle interact, and how the restricted lipid environment in these particles interferes with large scale conformational dynamics of the encased membrane protein. In a collaboration with Prof. Konstantin Shaitan, Lomonosov Moscow State University (Russia), we intend to scrutinize the properties of MACP-particles by combining experimental and theoretical approaches. Using different MACPs, we will characterize MACP-lipid/lipoprotein particles by size exclusion chromatography, dynamic light scattering, atomic-force microscopy, transmission electron microscopy, and EPR spectroscopy. As a functional detector the complex of the light sensitive archaeal sensory rhodopsin and its cognate transducer protein will be inserted into MACP-particles. Its protein dynamics will be followed on different time scales using time resolved optical and EPR spectroscopy to understand, how the nature of MACPs affects the light-induced signal transduction within the complex. Concurrently, our collaboration partner will model MACP-particles and analyze them by molecular dynamic simulations to assess how electrostatic, hydrophobic and stacking interactions stabilize the particles. The goals of the project are to draw a physico-chemical picture of formation and stabilization of MACP-encased lipid/lipoprotein nanoparticles and to develop optimized protocols of protein extraction by MACPs yielding native-like environments within MACP-lipid/lipoprotein particles.

含两亲性马来酸的交替共聚物（MACPs），其中马来酸单元与（可变）烃单元交替，是膜蛋白研究中最近的一个重大突破。发现几种MACP溶解来自人工和天然脂质双层的膜蛋白，产生直径在10-30 nm范围内均匀的盘状MACP包裹的脂质/脂蛋白颗粒。酶似乎在MACP颗粒内保持功能性;因此，这样的颗粒甚至使那些在去污剂存在下不稳定的膜蛋白可用于需要均质水溶性（单个）颗粒的技术。然而，它仍然是不清楚的，如何在没有洗涤剂的情况下，MACP提取膜蛋白，如何的MACP-脂质/脂蛋白颗粒的不同组分相互作用，以及如何在这些颗粒中的限制脂质环境干扰大规模的构象动力学的包裹的膜蛋白。在与罗蒙诺索夫莫斯科州立大学（俄罗斯）的康斯坦丁·谢坦教授的合作中，我们打算通过结合实验和理论方法来仔细研究MACP粒子的性质。 使用不同的MACP，我们将通过尺寸排阻色谱法，动态光散射，原子力显微镜，透射电子显微镜和EPR光谱表征MACP-脂质/脂蛋白颗粒。 作为功能检测器，光敏古生菌感觉视紫红质及其同源转导蛋白的复合物将被插入到MACP颗粒中。它的蛋白质动力学将使用时间分辨光学和EPR光谱在不同的时间尺度上进行跟踪，以了解MACPs的性质如何影响复合物内的光诱导信号转导。同时，我们的合作伙伴将对MACP颗粒进行建模，并通过分子动力学模拟对其进行分析，以评估静电，疏水和堆积相互作用如何稳定颗粒。该项目的目标是绘制形成和稳定的MACP包裹的脂质/脂蛋白纳米颗粒的物理化学图，并开发优化的协议，蛋白质提取的MACP内产生类似天然环境的MACP-脂质/脂蛋白颗粒。

项目成果

Evaluation of DIBMA nanoparticles of variable size and anionic lipid content as tools for the structural and functional study of membrane proteins.

不同尺寸和阴离子脂质含量的 DIBMA 纳米颗粒作为膜蛋白结构和功能研究工具的评估

• DOI: 10.1016/j.bbamem.2021.183588
• 发表时间: 2021
• 期刊: Biochimica et biophysica acta. Biomembranes
• 作者: Voskoboynikova N;Margheritis EG;Kodde F;Rademacher M;Schowe M;Budke-Gieseking A;Psathaki OE;Steinhoff HJ;Cosentino K
• 通讯作者: Cosentino K