Influence of DIBMA Polymer Length on Lipid Nanodisc Formation and Membrane Protein Extraction
Influence of DIBMA Polymer Length on Lipid Nanodisc Formation and Membrane Protein Extraction
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DOI：
10.1021/acs.biomac.0c01538
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发表时间：
2021-02-01
影响因子：
6.2
通讯作者：
Klumperman, Bert
中科院分区：
文献类型：
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作者：
Ball, Lauren E.;Riley, Liam J.;Klumperman, Bert
基于聚合物的脂质纳米颗粒，如苯乙烯马来酸脂质颗粒，彻底改变了膜蛋白的研究。最近，替代聚合物如聚（二异丁烯-交替-马来酸）（DIBMA）已用于该领域。DIBMA通常通过常规的自由基共聚来合成。为了研究其链长对脂质纳米盘形成和膜蛋白提取的影响，我们通过RAFT介导聚合合成了摩尔质量为1.2-12 kDa的DIBMA。对于3-7 kDa范围内的摩尔质量，脂质纳米盘形成的速率最高，并且与聚苯乙烯-共-马来酸（SMA）和市售DIBMA的速率相似。ZipA增溶效率显著高于市售DIBMA，与SMA相似（约75%）。此外，具有1.2- 3.9kDa的摩尔质量的RAFT制备的DIBMA在SDS-PAGE上显示出更干净的分离，而没有SMA和市售DIBMA通常所见的拖尾。
Polymer-based lipid nanoparticles like styrenemaleic acid lipid particles have revolutionized the study of membrane proteins. More recently, alternative polymers such as poly(diisobutylene-alt-maleic acid) (DIBMA) have been used in this field. DIBMA is commonly synthesized via conventional radical copolymerization. In order to study the influence of its chain length on lipid nanodisc formation and membrane protein extraction, we synthesized DIBMA with molar masses varying from 1.2-12 kDa via RAFT-mediated polymerization. For molar masses in the range of 3-7 kDa, the rate of lipid nanodisc formation was the highest and similar to those of polystyrene-co-maleic acid) (SMA) and commercially available DIBMA. ZipA solubilization efficiency was significantly higher than for commercially available DIBMA and similar to SMA (circa 75%). Furthermore, RAFT-made DIBMA with a molar mass of 1.2-3.9 kDa showed a much cleaner separation on SDS-PAGE, without the smearing that is typically seen for SMA and commercially available DIBMA.