Differences in SMA-like polymer architecture dictate the conformational changes exhibited by the membrane protein rhodopsin encapsulated in lipid nano-particles.
Differences in SMA-like polymer architecture dictate the conformational changes exhibited by the membrane protein rhodopsin encapsulated in lipid nano-particles.
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SMA类聚合物结构的不同决定了包裹在脂质纳米颗粒中的膜蛋白视紫红质的构象变化。
DOI：
10.1039/d1nr02419a
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发表时间：
2021-08-21
期刊：
影响因子：
6.7
通讯作者：
Wheatley M
中科院分区：
文献类型：
--
作者：
Grime RL ;Logan RT ;Nestorow SA ;Sridhar P ;Edwards PC ;Tate CG ;Klumperman B ;Dafforn TR ;Poyner DR ;Reeves PJ ;Wheatley M
膜蛋白在细胞过程中起着至关重要的作用，而保持其天然脂质双层环境的纳米包囊策略对于研究和开发这些蛋白特别有吸引力。聚(苯乙烯-马来酸)(SMA)及其相关聚合物聚(苯乙烯-co-(N-(3-N‘，N’-二甲氨基丙基)马来酰亚胺)(SMI)和聚(二异丁烯-ALT-马来酸)(DIBMA)通过自发地直接从天然双层的纳米盘中溶解膜蛋白，分别称为SMA脂质颗粒(SMALPs)、SMILPs和DIBMALPs)，已经给膜蛋白的研究带来了革命性的变化。这项系统的研究首次表明，被包裹的蛋白质的构象变化是由增溶聚合物决定的。视紫红质(Rho)是一种G蛋白偶联受体(GPCR)，它的光激活途径由结构定义的中间体组成，具有特征吸收光谱，显示了与含苯乙烯的SMA和SMI的构象限制，因此在SMALP或SMILP中，光激活仅进行到478 nm吸收的变视紫红质-I。相反，在DIBMALP中观察到了在382 nm处吸收的变视紫红质-II的完全达到。因此，通过简单地使用不同的SMA类聚合物，可以容易地产生不同的RHO中间态。动态光散射和分析超速离心法发现SMALP、SMILP和DIBMALP在尺寸和热稳定性方面存在差异。此外，包裹的Rho在SMALP、SMILP和DIBMALP中表现出不同的稳定性。总之，我们确定SMA、SMI和DIBMA构成了增溶聚合物的“工具箱”，因此选择合适的增溶聚合物为研究膜蛋白提供了一系列有用的属性。以GPCR视紫质为例，SMA、SMI和DIBMA构成了结构相关增溶聚合物的“工具包”，每一种都为研究包裹在脂质颗粒中的膜蛋白提供了不同的优势。
Membrane proteins are of fundamental importance to cellular processes and nano-encapsulation strategies that preserve their native lipid bilayer environment are particularly attractive for studying and exploiting these proteins. Poly(styrene-co-maleic acid) (SMA) and related polymers poly(styrene-co-(N-(3-N′,N′-dimethylaminopropyl)maleimide)) (SMI) and poly(diisobutylene-alt-maleic acid) (DIBMA) have revolutionised the study of membrane proteins by spontaneously solubilising membrane proteins direct from cell membranes within nanoscale discs of native bilayer called SMA lipid particles (SMALPs), SMILPs and DIBMALPs respectively. This systematic study shows for the first time, that conformational changes of the encapsulated protein are dictated by the solubilising polymer. The photoactivation pathway of rhodopsin (Rho), a G-protein-coupled receptor (GPCR), comprises structurally-defined intermediates with characteristic absorbance spectra that revealed conformational restrictions with styrene-containing SMA and SMI, so that photoactivation proceeded only as far as metarhodopsin-I, absorbing at 478 nm, in a SMALP or SMILP. In contrast, full attainment of metarhodopsin-II, absorbing at 382 nm, was observed in a DIBMALP. Consequently, different intermediate states of Rho could be generated readily by simply employing different SMA-like polymers. Dynamic light-scattering and analytical ultracentrifugation revealed differences in size and thermostability between SMALP, SMILP and DIBMALP. Moreover, encapsulated Rho exhibited different stability in a SMALP, SMILP or DIBMALP. Overall, we establish that SMA, SMI and DIBMA constitute a ‘toolkit’ of solubilising polymers, so that selection of the appropriate solubilising polymer provides a spectrum of useful attributes for studying membrane proteins. Using the GPCR rhodopsin as an exemplar, SMA SMI and DIBMA constitute a ‘tool-kit’ of structurally-related solubilising polymers, with each providing different advantages for studying membrane proteins encapsulated in lipid particles.
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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
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Wheatley M
影响因子：
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影响因子：
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作者：
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通讯作者：
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影响因子：
3.4
作者：
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通讯作者：
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使用二异丁基马来酸（DIBMA）共聚物对功能性人类5-羟色胺转运蛋白（SERT）的无洗涤剂纯化和重构。
DOI：
10.1016/j.bbamem.2021.183602
发表时间：
2021-07-01
期刊：
Biochimica et biophysica acta. Biomembranes
影响因子：
--
作者：
Dilworth MV;Findlay HE;Booth PJ
通讯作者：
Booth PJ