Expression, Purification, and Structure Determination of Human PTCH1-HH-N Complexes.
Expression, Purification, and Structure Determination of Human PTCH1-HH-N Complexes.
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人 PTCH1-HH-N 复合物的表达、纯化和结构测定。
DOI：
10.1007/978-1-0716-1701-4_10
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Li,Xiaochun
中科院分区：
文献类型：
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作者：
Qi,Xiaofeng;Schmiege,Philip;Esparza,Leticia;Li,Xiaochun
Patched-1（PTCH 1）是一种肿瘤抑制因子，作为Hedgehog（HH）配体的受体，负性调节HH信号通路。PTCH 1的突变与许多人类癌症有关。结构研究揭示了PTCH 1介导的HH信号调节机制，进一步促进了癌症的治疗发展。在这里，我们描述了一个几乎全长的功能PTCH 1变体，PTCH 1 * 的表达和纯化。用纯化的PTCH 1 * 蛋白组装了两种形式的PTCH 1 *-Sonic Hedgehog（SHH）复合物，并随后通过冷冻电镜（cryo-EM）确定了它们的结构。
Patched-1 (PTCH1), a tumor suppressor, serves as the receptor of Hedgehog (HH) ligand and negatively regulates the HH signaling pathway. Mutations of PTCH1 are implicated in many human cancers. Structural investigation revealed the mechanism of PTCH1-mediated HH signal regulation, further facilitating the therapeutic development of cancers. Here, we describe the expression and purification of a nearly full-length functional PTCH1 variant, PTCH1*. With purified PTCH1* protein, two forms of PTCH1*–Sonic Hedgehog (SHH) complexes were assembled, and their structures subsequently determined by cryo-electron microscope (cryo-EM).