Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
Eur Biophys J. 2023 Feb;52(1-2):39-51. doi: 10.1007/s00249-023-01632-5. Epub 2023 Feb 14.
From the discovery of the first membrane-interacting polymer, styrene maleic-acid (SMA), there has been a rapid development of membrane solubilising polymers. These new polymers can solubilise membranes under a wide range of conditions and produce varied sizes of nanoparticles, yet there has been a lack of broad comparison between the common polymer types and solubilising conditions. Here, we present a comparative study on the three most common commercial polymers: SMA 3:1, SMA 2:1, and DIBMA. Additionally, this work presents, for the first time, a comparative characterisation of polymethacrylate copolymer (PMA). Absorbance and dynamic light scattering measurements were used to evaluate solubilisation across key buffer conditions in a simple, adaptable assay format that looked at pH, salinity, and divalent cation concentration. Lipid-polymer nanoparticles formed from SMA variants were found to be the most susceptible to buffer effects, with nanoparticles from either zwitterionic DMPC or POPC:POPG (3:1) bilayers only forming in low to moderate salinity (< 600 mM NaCl) and above pH 6. DIBMA-lipid nanoparticles could be formed above a pH of 5 and were stable in up to 4 M NaCl. Similarly, PMA-lipid nanoparticles were stable in all NaCl concentrations tested (up to 4 M) and a broad pH range (3-10). However, for both DIBMA and PMA nanoparticles there is a severe penalty observed for bilayer solubilisation in non-optimal conditions or when using a charged membrane. Additionally, lipid fluidity of the DMPC-polymer nanoparticles was analysed through cw-EPR, showing no cooperative gel-fluid transition as would be expected for native-like lipid membranes.
从发现第一种与膜相互作用的聚合物马来酸-苯乙烯(SMA)以来,用于溶解膜的聚合物得到了迅速发展。这些新的聚合物可以在广泛的条件下溶解膜,并产生不同大小的纳米颗粒,但对于常见的聚合物类型和溶解条件,缺乏广泛的比较。在这里,我们对三种最常见的商业聚合物:SMA 3:1、SMA 2:1 和 DIBMA 进行了比较研究。此外,这项工作首次对聚甲基丙烯酸甲酯共聚物(PMA)进行了比较研究。吸光度和动态光散射测量用于评估在简单、适应性强的测定格式中,关键缓冲条件下的溶解情况,该格式研究了 pH 值、盐度和二价阳离子浓度。SMA 变体形成的脂质-聚合物纳米颗粒最容易受到缓冲液的影响,来自两性离子 DMPC 或 POPC:POPG(3:1)双层的纳米颗粒仅在低盐度(<600 mM NaCl)和 pH 值高于 6 时形成。DIBMA-脂质纳米颗粒可以在 pH 值高于 5 且在高达 4 M NaCl 下稳定。同样,PMA-脂质纳米颗粒在所有测试的 NaCl 浓度(高达 4 M)和宽 pH 范围内(3-10)都是稳定的。然而,对于 DIBMA 和 PMA 纳米颗粒,在非最佳条件下或使用带电膜时,双层的溶解都会出现严重的损失。此外,通过连续波 EPR 分析了 DMPC-聚合物纳米颗粒的脂质流动性,没有观察到类似天然脂质膜的协同凝胶-流态转变。