An acid-compatible co-polymer for the solubilization of membranes and proteins into lipid bilayer-containing nanoparticles.
An acid-compatible co-polymer for the solubilization of membranes and proteins into lipid bilayer-containing nanoparticles.
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DOI：
10.1039/c8nr01322e
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发表时间：
2018-06-07
期刊：
影响因子：
6.7
通讯作者：
Dafforn TR
中科院分区：
文献类型：
--
作者：
Hall SCL ;Tognoloni C ;Charlton J ;Bragginton ÉC ;Rothnie AJ ;Sridhar P ;Wheatley M ;Knowles TJ ;Arnold T ;Edler KJ ;Dafforn TR
带正电的聚(苯乙烯-马来酰亚胺)将功能性膜蛋白提取到纳米盘中，克服了目前纳米盘技术的一些局限性。膜蛋白在药物开发中的重要作用意味着膜模拟系统在膜蛋白研究中的应用越来越受到人们的关注。一种这样的体系是两亲性的、带负电荷的聚(苯乙烯-马来酸)(SMA)聚合物，形成“SMA类脂颗粒”(SMALPs)，它已被广泛采用来直接从细胞膜中溶解膜蛋白。然而，SMALP只有在碱性条件下才能溶解，并在许多下游应用所需的二价阳离子存在下沉淀。在这里，我们展示了带正电的聚(苯乙烯-马来酰亚胺)(SMI)形成类似的纳米颗粒，具有与SMA相同的效率，同时在酸性pH下保持功能，并与高浓度的二价阳离子兼容。我们对SMI的性能进行了详细的表征，使其能够与为SMA发布的类似数据进行直接比较。我们还证明了SMI能够直接从细胞膜中提取蛋白质，并能够溶解在培养的HEK 293T细胞中表达的功能性人G蛋白偶联受体(GPCRs)。因此，“SMILP”提供了一种替代的膜增溶方法，它成功地克服了SMALP方法的一些局限性。
Positively charged poly(styrene-co-maleimide) extracts functional membrane proteins into nanodiscs, overcoming some limitations of current nanodisc technology. The fundamental importance of membrane proteins in drug discovery has meant that membrane mimetic systems for studying membrane proteins are of increasing interest. One such system has been the amphipathic, negatively charged poly(styrene-co-maleic acid) (SMA) polymer to form “SMA Lipid Particles” (SMALPs) which have been widely adopted to solubilize membrane proteins directly from the cell membrane. However, SMALPs are only soluble under basic conditions and precipitate in the presence of divalent cations required for many downstream applications. Here, we show that the positively charged poly(styrene-co-maleimide) (SMI) forms similar nanoparticles with comparable efficiency to SMA, whilst remaining functional at acidic pH and compatible with high concentrations of divalent cations. We have performed a detailed characterization of the performance of SMI that enables a direct comparison with similar data published for SMA. We also demonstrate that SMI is capable of extracting proteins directly from the cell membrane and can solubilize functional human G-protein coupled receptors (GPCRs) expressed in cultured HEK 293T cells. “SMILPs” thus provide an alternative membrane solubilization method that successfully overcomes some of the limitations of the SMALP method.
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影响因子：
4
作者：
Jamshad M;Charlton J;Lin YP;Routledge SJ;Bawa Z;Knowles TJ;Overduin M;Dekker N;Dafforn TR;Bill RM;Poyner DR;Wheatley M
通讯作者：
Wheatley M
影响因子：
5.4
作者：
Almen, Markus Sallman;Nordstrom, Karl J. V.;Schioth, Helgi B.
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影响因子：
3.4
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通讯作者：
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影响因子：
4.8
作者：
Mishra, Vinod K.;Palgunachari, Mayakonda N.;Anantharamaiah, G. M.
通讯作者：
Anantharamaiah, G. M.
影响因子：
3.4
作者：
Midtgaard, Soren Roi;Pedersen, Martin Cramer;Arleth, Lise
通讯作者：
Arleth, Lise