Membrane protein extraction and purification using partially-esterified SMA polymers.
Membrane protein extraction and purification using partially-esterified SMA polymers.
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DOI：
10.1016/j.bbamem.2021.183758
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发表时间：
2021-12-01
期刊：
影响因子：
--
通讯作者：
Rothnie AJ
中科院分区：
文献类型：
--
作者：
Hawkins OP;Jahromi CPT;Gulamhussein AA;Nestorow S;Bahra T;Shelton C;Owusu-Mensah QK;Mohiddin N;O'Rourke H;Ajmal M;Byrnes K;Khan M;Nahar NN;Lim A;Harris C;Healy H;Hasan SW;Ahmed A;Evans L;Vaitsopoulou A;Akram A;Williams C;Binding J;Thandi RK;Joby A;Guest A;Tariq MZ;Rasool F;Cavanagh L;Kang S;Asparuhov B;Jestin A;Dafforn TR;Simms J;Bill RM;Goddard AD;Rothnie AJ
苯乙烯马来酸（SMA）聚合物已被证明是非常成功的膜蛋白提取，形成SMA脂质颗粒（SMALPs），它在膜蛋白周围维持了一层脂质双分子层。small - p包封膜蛋白可用于功能和结构研究。small方法允许保留重要的蛋白质-环状脂质相互作用，施加侧压力，并提供比传统洗涤剂增溶更大的稳定性。然而，SMA聚合物确实有一些局限性，包括对二价阳离子和低pH值的敏感性，与许多蛋白质重叠的吸光度光谱，以及对蛋白质构象变化的可能限制。人们开发了各种改性聚合物，试图克服这些挑战，但还没有找到明确的解决方案。SMA的一系列部分酯化变体（SMA 2625， SMA 1440和SMA 17352）先前已被证明对植物和蓝藻类囊体膜的增溶非常有效。假设马来酸基团的部分酯化会增加对二价阳离子的耐受性。因此，我们测试了这些部分酯化的聚合物对脂质和膜蛋白的增溶性，以及它们对镁离子的耐受性。结果表明，所有部分酯化聚合物都能增溶和纯化一系列膜蛋白，但SMA 1440的蛋白收率较低，SMA 1440和SMA 17352的纯度都较低。SMA 2625的表现与SMA 2000相当。SMA 1440也显示出对二价阳离子的敏感性增加。因此，SMA和二价阳离子之间的相互作用似乎比提出的更复杂，需要进一步研究。部分酯化的SMA聚合物对于不同表达系统的膜蛋白的溶解是有效的。部分酯化的SMALPs蛋白可以用Ni2+亲和层析纯化，但用SMA 1440纯化得率较低。SMA的部分酯化不能克服对二价阳离子的敏感性。SMA1440对镁离子的敏感性高于sma2000。
Styrene maleic acid (SMA) polymers have proven to be very successful for the extraction of membrane proteins, forming SMA lipid particles (SMALPs), which maintain a lipid bilayer around the membrane protein. SMALP-encapsulated membrane proteins can be used for functional and structural studies. The SMALP approach allows retention of important protein-annular lipid interactions, exerts lateral pressure, and offers greater stability than traditional detergent solubilisation. However, SMA polymer does have some limitations, including a sensitivity to divalent cations and low pH, an absorbance spectrum that overlaps with many proteins, and possible restrictions on protein conformational change. Various modified polymers have been developed to try to overcome these challenges, but no clear solution has been found. A series of partially-esterified variants of SMA (SMA 2625, SMA 1440 and SMA 17352) has previously been shown to be highly effective for solubilisation of plant and cyanobacterial thylakoid membranes. It was hypothesised that the partial esterification of maleic acid groups would increase tolerance to divalent cations. Therefore, these partially-esterified polymers were tested for the solubilisation of lipids and membrane proteins, and their tolerance to magnesium ions. It was found that all partially esterified polymers were capable of solubilising and purifying a range of membrane proteins, but the yield of protein was lower with SMA 1440, and the degree of purity was lower for both SMA 1440 and SMA 17352. SMA 2625 performed comparably to SMA 2000. SMA 1440 also showed an increased sensitivity to divalent cations. Thus, it appears the interactions between SMA and divalent cations are more complex than proposed and require further investigation. Partially-esterified SMA polymers are effective for solubilisation of membrane proteins from different expression systems. Proteins in partially-esterified SMALPs can be purified by Ni2+ affinity chromatography, but yield is lower with SMA 1440. Partial esterification of SMA does not overcome the sensitivity to divalent cations. SMA1440 is more sensitive to magnesium ions than SMA 2000.
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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：
6.7
作者：
Hall SCL ;Tognoloni C ;Charlton J ;Bragginton ÉC ;Rothnie AJ ;Sridhar P ;Wheatley M ;Knowles TJ ;Arnold T ;Edler KJ ;Dafforn TR
通讯作者：
Dafforn TR
影响因子：
3.9
作者：
通讯作者：
--
影响因子：
3.3
作者：
Kherb, Jaibir;Flores, Sarah C.;Cremer, Paul S.
通讯作者：
Cremer, Paul S.
影响因子：
3.4
作者：
Kopf, Adrian H.;Dorr, Jonas M.;Killian, J. Antoinette
通讯作者：
Killian, J. Antoinette