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

Evaluation of DIBMA nanoparticles of variable size and anionic lipid content as tools for the structural and functional study of membrane proteins.
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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
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发表时间:
2021
期刊:
Biochimica et biophysica acta. Biomembranes
影响因子:
--
通讯作者:
Cosentino K
Cosentino K
中科院分区:
--
文献类型:
--
作者:
Voskoboynikova N;Margheritis EG;Kodde F;Rademacher M;Schowe M;Budke-Gieseking A;Psathaki OE;Steinhoff HJ;Cosentino K

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含两亲性马来酸的聚合物允许将膜蛋白直接提取成稳定的、均质的、水溶性共聚物/脂质纳米颗粒,而不使用洗涤剂。通过调节聚合物/脂质的比例,可以方便地调节纳米颗粒的尺寸,以掺入不同尺寸的蛋白质复合物。然而,脂质纳米颗粒尺寸的增加可能与样品异质性的增加相关,从而阻碍了它们在需要高度均匀样品的光谱和结构技术中的应用。此外,尺寸均匀性可以受到影响的低脂质体溶解效率的DIBMA,它携带负电荷,在存在高的脂质电荷density.In这项工作中,我们应用生物物理工具来表征的大小和大小的异质性大(15 nm以上)的脂质纳米颗粒包裹的二异丁烯/马来酸(DIBMA)共聚物在不同的DIBMA/脂质的比例和阴离子脂质的百分比。重要的是,对于直径范围为40 nm或更小的纳米颗粒制剂,DIBMA/脂质纳米颗粒(DIBMALP)的尺寸均匀性保持不变。此外,我们表明,阴离子脂质不影响DIBMALP的生产,大小和大小均匀性。此外,它们不影响膜中的整体脂质动力学,并保留了封闭的膜蛋白的功能性,这项工作加强了DIBMALP作为通用的,天然的膜蛋白的功能研究的脂质环境的适用性,并提供有用的洞察力,这些系统的适用性,这些结构的技术,需要高度均匀的样品制备。
Amphiphilic maleic acid-containing polymers allow for the direct extraction of membrane proteins into stable, homogenous, water-soluble copolymer/lipid nanoparticles without the use of detergents. By adjusting the polymer/lipid ratio, the size of the nanoparticles can be tuned at convenience for the incorporation of protein complexes of different size. However, an increase in the size of the lipid nanoparticles may correlate with increased sample heterogeneity, thus hampering their application to spectroscopic and structural techniques where highly homogeneous samples are desirable. In addition, size homogeneity can be affected by low liposome solubilization efficiency by DIBMA, which carries a negative charge, in the presence of high lipid charge density.In this work, we apply biophysical tools to characterize the size and size heterogeneity of large (above 15 nm) lipid nanoparticles encased by the diisobutylene/maleic acid (DIBMA) copolymer at different DIBMA/lipid ratios and percentages of anionic lipids. Importantly, for nanoparticle preparations in the diameter range of 40 nm or below, the size homogeneity of the DIBMA/lipid nanoparticles (DIBMALPs) remains unchanged. In addition, we show that anionic lipids do not affect the production, size and size homogeneity of DIBMALPs. Furthermore, they do not affect the overall lipid dynamics in the membrane, and preserve the functionality of an enclosed membrane protein.This work strengthens the suitability of DIBMALPs as universal, native-like lipid environments for functional studies of membrane proteins and provide useful insight on the suitability of these systems for those structural techniques requiring highly homogeneous sample preparations.
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