Department of Physiology and Cellular Biophysics, Columbia University, New York, NY, USA.
J Struct Biol. 2010 Oct;172(1):85-93. doi: 10.1016/j.jsb.2010.03.021. Epub 2010 Apr 13.
High-throughput (HT) methodologies have had a tremendous impact on structural biology of soluble proteins. High-resolution structure determination relies on the ability of the macromolecule to form ordered crystals that diffract X-rays. While crystallization remains somewhat empirical, for a given protein, success is proportional to the number of conditions screened and to the number of variants trialed. HT techniques have greatly increased the number of targets that can be trialed and the rate at which these can be produced. In terms of number of structures solved, membrane proteins appear to be lagging many years behind their soluble counterparts. Likewise, HT methodologies for production and characterization of these hydrophobic macromolecules are only now emerging. Presented here is an HT platform designed exclusively for membrane proteins that has processed over 5000 targets.
高通量 (HT) 方法极大地推动了可溶性蛋白质的结构生物学研究。高分辨率结构测定依赖于大分子形成有序晶体从而使 X 射线产生衍射的能力。尽管结晶仍然具有一定的经验性,但对于给定的蛋白质,成功与否与筛选的条件数量和尝试的变体数量成正比。HT 技术大大增加了可以尝试的目标数量以及这些目标的生产速度。就解决的结构数量而言,膜蛋白似乎比其可溶性对应物落后了许多年。同样,用于这些疏水分子的生产和特性描述的 HT 方法现在才刚刚出现。本文介绍了一个专门为膜蛋白设计的 HT 平台,该平台已经处理了超过 5000 个目标。