@inbook{6ed9107be2d64d98810b0d2051f20977,
title = "Expression, Purification, and Structure Determination of Human PTCH1–HH-N Complexes",
abstract = "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).",
keywords = "Complex assembly, Cryo-EM, Hedgehog, Patched-1, Protein purification",
author = "Xiaofeng Qi and Philip Schmiege and Leticia Esparza and Xiaochun Li",
note = "Funding Information: This work was supported by the Endowed Scholars Program in Medical Science of UT Southwestern Medical Center and NIH grant R01 GM135343 (to X.L.). X.Q. is the recipient of a DDBrown Fellow of the Life Sciences Research Foundation. P.S. is supported by NIH grant T32 GM131963. X.L. is a Damon Runyon-Rachleff Innovator supported by the Damon Runyon Cancer Research Foundation (DRR-53-19) and a Rita C. and William P. Clements, Jr. Scholar in Biomedical Research at UT Southwestern Medical Center. Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-1701-4_10",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "107--120",
booktitle = "Methods in Molecular Biology",
}