Yeast as a tool for membrane protein production and structure determination.
Yeast as a tool for membrane protein production and structure determination.
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DOI：
10.1093/femsyr/foac047
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发表时间：
2022-10-20
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
--
作者：
膜蛋白是功能和结构表征具有挑战性的目标。他们研究中的一个持久瓶颈是可靠地生产足够产量的稳定蛋白质。在这里，我们评估了所有支持高分辨率结构沉积的真核膜蛋白生产实验。我们专注于最常见的酵母宿主系统，酿酒酵母和毕赤酵母。 1999 年首次描述了酵母中产生的膜蛋白的高分辨率结构，如今有 186 种 α-螺旋膜蛋白结构，代表来自 37 个家族的 101 种独特蛋白质。同源和异源生产在酿酒酵母中同样常见，而异源生产在毕赤酵母中占主导地位，尤其是人类蛋白质，约占总数的三分之一。研究蛋白质工程方法（来自七个家族的 78 种蛋白质）表明，大多数都包含用于纯化的多组氨酸标签，通常位于蛋白质的 C 末端。密码子优化和亲水性延伸截断也是提高产量的常见方法。我们得出的结论是，酵母仍然是研究 α-螺旋膜蛋白的有用生产宿主。尽管大多数真核膜蛋白结构源自 HEK293 和昆虫细胞中产生的蛋白质，但作者在此展示了酵母作为生产宿主的重要性及其作为用于结构和功能分析的真核膜蛋白生产中的重要参与者的作用。
Membrane proteins are challenging targets to functionally and structurally characterize. An enduring bottleneck in their study is the reliable production of sufficient yields of stable protein. Here, we evaluate all eukaryotic membrane protein production experiments that have supported the deposition of a high-resolution structure. We focused on the most common yeast host systems, Saccharomyces cerevisiae and Pichia pastoris. The first high-resolution structure of a membrane protein produced in yeast was described in 1999 and today there are 186 structures of α-helical membrane proteins, representing 101 unique proteins from 37 families. Homologous and heterologous production are equally common in S. cerevisiae, while heterologous production dominates in P. pastoris, especially of human proteins, which represent about one-third of the total. Investigating protein engineering approaches (78 proteins from seven families) demonstrated that the majority contained a polyhistidine tag for purification, typically at the C-terminus of the protein. Codon optimization and truncation of hydrophilic extensions were also common approaches to improve yields. We conclude that yeast remains a useful production host for the study of α-helical membrane proteins. Although the majority of eukaryotic MEMBRANE PROTEIN structures are DERIVED FROM PROTEINS produced in HEK293 and insect cells, the authors show here the importance of yeast as a production host and its role as an essential player in the production of eukaryotic membrane proteins for structural and functional analysis.
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影响因子：
3.4
作者：
Bill，Roslyn M.;Hedfalk，Kristina
通讯作者：
Hedfalk，Kristina
影响因子：
5.8
作者：
Robinson GC;Bason JV;Montgomery MG;Fearnley IM;Mueller DM;Leslie AG;Walker JE
通讯作者：
Walker JE
影响因子：
11.4
作者：
Clausen, Johannes D.;Bublitz, Maike;le Maire, Marc
通讯作者：
le Maire, Marc
DOI：
10.1126/science.1168750
发表时间：
2009-03-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Aller SG;Yu J;Ward A;Weng Y;Chittaboina S;Zhuo R;Harrell PM;Trinh YT;Zhang Q;Urbatsch IL;Chang G
通讯作者：
Chang G
影响因子：
16.6
作者：
Bai L;Jain BK;You Q;Duan HD;Takar M;Graham TR;Li H
通讯作者：
Li H