Loll Patrick J
Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, 245 North 15th Street, Philadelphia, PA 19102, USA.
Acta Crystallogr F Struct Biol Commun. 2014 Dec 1;70(Pt 12):1576-83. doi: 10.1107/S2053230X14025035. Epub 2014 Nov 28.
At the time when the first membrane-protein crystal structure was determined, crystallization of these molecules was widely perceived as extremely arduous. Today, that perception has changed drastically, and the process is regarded as routine (or nearly so). On the occasion of the International Year of Crystallography 2014, this review presents a snapshot of the current state of the art, with an emphasis on the role of detergents in this process. A survey of membrane-protein crystal structures published since 2012 reveals that the direct crystallization of protein-detergent complexes remains the dominant methodology; in addition, lipidic mesophases have proven immensely useful, particularly in specific niches, and bicelles, while perhaps undervalued, have provided important contributions as well. Evolving trends include the addition of lipids to protein-detergent complexes and the gradual incorporation of new detergents into the standard repertoire. Stability has emerged as a critical parameter controlling how a membrane protein behaves in the presence of detergent, and efforts to enhance stability are discussed. Finally, although discovery-based screening approaches continue to dwarf mechanistic efforts to unravel crystallization, recent technical advances offer hope that future experiments might incorporate the rational manipulation of crystallization behaviors.
在首个膜蛋白晶体结构被确定之时,这些分子的结晶被广泛认为极其艰巨。如今,这种看法已发生巨大改变,该过程被视为常规操作(或几乎如此)。值此2014年国际晶体学年之际,本综述呈现了当前技术水平的概况,重点关注去污剂在这一过程中的作用。对2012年以来发表的膜蛋白晶体结构的调查表明,蛋白质 - 去污剂复合物的直接结晶仍然是主要方法;此外,脂质中间相已证明非常有用,特别是在特定领域,而双分子层虽然可能未得到充分重视,但也做出了重要贡献。不断发展的趋势包括在蛋白质 - 去污剂复合物中添加脂质以及新的去污剂逐渐纳入标准范围。稳定性已成为控制膜蛋白在去污剂存在下行为的关键参数,并讨论了增强稳定性的努力。最后,尽管基于发现的筛选方法在解开结晶的机理研究方面仍占主导地位,但最近的技术进展带来了希望,即未来的实验可能会纳入对结晶行为的合理操控。