Expression, Purification, and Structure Determination of Human PTCH1–HH-N Complexes

Xiaofeng Qi, Philip Schmiege, Leticia Esparza, Xiaochun Li

Research output: Chapter in Book/Report/Conference proceedingChapter


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).

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages14
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Complex assembly
  • Cryo-EM
  • Hedgehog
  • Patched-1
  • Protein purification

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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