Obtaining Crystals of PPARγ Ligand Binding Domain Bound to Small Molecules
Obtaining Crystals of PPARγ Ligand Binding Domain Bound to Small Molecules
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DOI：
10.1007/978-1-4939-9195-2_21
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发表时间：
2019-01-01
期刊：
影响因子：
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通讯作者：
Bruning, John B.
中科院分区：
文献类型：
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作者：
Frkic, Rebecca L.;Bruning, John B.
获得与其蛋白质靶点结合的小分子候选药物的高通量结构表征受到共结晶蛋白质-配体复合物的挑战的阻碍。这些挑战包括配体溶解性差、过量的配体分子破坏待结晶样品的均匀性、以及在为每种候选药物制备单独的复合物和晶体筛选中的效率低下。将载脂蛋白结晶，然后用含有目标配体的溶液浸泡，是快速获得配体结构信息的有力工具。在这里，我们描述了纯化，结晶和浸泡的过程中的过氧化物酶体增殖物激活受体。配体结合结构域以及用于共结晶不适于浸泡的配体的策略。
Obtaining high-throughput structural characterization of small molecule drug candidates bound to their protein target has been hampered by the challenges of cocrystallizing protein-ligand complexes. These challenges include poor ligand solubility, excess ligand molecules disrupting the homogeneity of the sample to be crystallized, and inefficiency in preparing individual complexes and crystal screens for each drug candidate. Crystallizing apo protein followed by soaking with a solution containing the ligand of interest is a powerful tool for rapidly obtaining structural information of ligands. Here, we describe the process of purifying, crystallizing, and soaking PPAR. ligand binding domain as well as strategies for cocrystallizing ligands that are not amenable to soaking.