Poly(styrene-co-maleic acid)-mediated isolation of supramolecular membrane protein complexes from plant thylakoids
Poly(styrene-co-maleic acid)-mediated isolation of supramolecular membrane protein complexes from plant thylakoids
复制标题
DOI：
10.1016/j.bbabio.2020.148347
复制
发表时间：
2021-03-01
影响因子：
4.3
通讯作者：
Bruce, Barry D.
中科院分区：
文献类型：
--
作者：
Korotych, Olena, I;Nguyen, Thao T.;Bruce, Barry D.
聚（苯乙烯-共-马来酸）（pSMA）的衍生物，最近出现作为从生物膜中提取膜蛋白复合物的有效试剂。尽管最近在使用SMA研究人工和细菌膜方面取得了进展，但很少有报道涉及它们在研究高度丰富和充分表征的类囊体膜中的用途。最近，我们测试了12个市售的SMA共聚物具有不同的物理化学性质，从菠菜类囊体膜提取膜蛋白复合物（MPC）的能力。基于蛋白质和叶绿素提取的功效，我们已经发现了五种高效SMA共聚物：SMA（R）1440、XIRAN（R）25010、XIRAN（R）30010、SMA（R）17352和SMA（R）PRO 10235，其显示出从叶绿体类囊体提取MPC的前景。为了进一步推进这些聚合物用于研究生物膜组织的应用，我们研究了由上述五种SMA聚合物提取的类囊体超分子蛋白质复合物的组成。两种常用的研究植物，菠菜（菠菜）和豌豆（豌豆），用于提取模型生物膜。我们发现，pSMA差异提取蛋白质复合物从菠菜和豌豆类囊体。基于其与聚合物化学结构相关的不同活性，pSMA可分为两组：未官能化的聚合物和酯衍生物。
Derivatives of poly(styrene-co-maleic acid) (pSMA), have recently emerged as effective reagents for extracting membrane protein complexes from biological membranes. Despite recent progress in using SMAs to study artificial and bacterial membranes, very few reports have addressed their use in studying the highly abundant and well characterized thylakoid membranes. Recently, we tested the ability of twelve commercially available SMA copolymers with different physicochemical properties to extract membrane protein complexes (MPCs) from spinach thylakoid membrane. Based on the efficacy of both protein and chlorophyll extraction, we have found five highly efficient SMA copolymers: SMA (R) 1440, XIRAN (R) 25010, XIRAN (R) 30010, SMA (R) 17352, and SMA (R) PRO 10235, that show promise in extracting MPCs from chloroplast thylakoids. To further advance the application of these polymers for studying biomembrane organization, we have examined the composition of thylakoid supramolecular protein complexes extracted by the five SMA polymers mentioned above. Two commonly studied plants, spinach (Spinacia oleracea) and pea (Pisum sativum), were used for extraction as model biomembranes. We found that the pSMAs differentially extract protein complexes from spinach and pea thylakoids. Based on their differential activity, which correlates with the polymer chemical structure, pSMAs can be divided into two groups: unfunctionalized polymers and ester derivatives.