Department of Health Chemistry, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
Faculty of Life and Environmental Sciences, University of Yamanashi, Kofu, Yamanashi 400-8510, Japan.
STAR Protoc. 2021 Feb 26;2(1):100364. doi: 10.1016/j.xpro.2021.100364. eCollection 2021 Mar 19.
Peroxisome proliferator-activated receptors (PPARs) are nuclear receptor-type transcription factors with three subtypes (α, δ, and γ) that regulate cell differentiation and metabolism. Co-crystals of human PPARα-ligand-binding domain (LBD)-PPARα ligand for X-ray crystallography have been difficult to obtain. Recombinant human PPARα-LBD proteins contain intrinsic fatty acids (iFAs of origin) and may be unstable without ligands during crystallization. To circumvent these limitations, we have successfully applied various crystallization techniques, including co-crystallization, cross-seeding, soaking, delipidation, and coactivator peptide supplementation. For complete details on the use and execution of this protocol, please refer to Kamata et al. (2020).
过氧化物酶体增殖物激活受体 (PPARs) 是核受体型转录因子，有三个亚型 (α、δ 和 γ)，可调节细胞分化和代谢。用于 X 射线晶体学的人 PPARα-配体结合域 (LBD)-PPARα配体的共结晶一直难以获得。重组人 PPARα-LBD 蛋白含有内源性脂肪酸（源自 iFA），在结晶过程中没有配体时可能不稳定。为了克服这些限制，我们已经成功应用了各种结晶技术，包括共结晶、交叉接种、浸泡、去脂和辅激活肽补充。有关此方案使用和执行的完整详细信息，请参阅 Kamata 等人。(2020)。
