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PTCHD1 Binds Cholesterol but Not Sonic Hedgehog, Suggesting a Distinct Cellular Function.
Hiltunen, Mimmu K; Timmis, Alex J; Thomsen, Maren; Gkotsi, Danai S; Iwaï, Hideo; Ribeiro, Orquidea M; Goldman, Adrian; Riobo-Del Galdo, Natalia A.
Afiliação
  • Hiltunen MK; Faculty of Biological and Environmental Sciences, University of Helsinki, 00100 Helsinki, Finland.
  • Timmis AJ; School of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Thomsen M; Astbury Center for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Gkotsi DS; School of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Iwaï H; Astbury Center for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Ribeiro OM; Institute of Biotechnology, HiLiFE, University of Helsinki, 00100 Helsinki, Finland.
  • Goldman A; Faculty of Biological and Environmental Sciences, University of Helsinki, 00100 Helsinki, Finland.
  • Riobo-Del Galdo NA; Faculty of Biological and Environmental Sciences, University of Helsinki, 00100 Helsinki, Finland.
Int J Mol Sci ; 24(3)2023 Jan 31.
Article em En | MEDLINE | ID: mdl-36769003
Deleterious mutations in the X-linked Patched domain-containing 1 (PTCHD1) gene may account for up to 1% of autism cases. Despite this, the PTCHD1 protein remains poorly understood. Structural similarities to Patched family proteins point to a role in sterol transport, but this hypothesis has not been verified experimentally. Additionally, PTCHD1 has been suggested to be involved in Hedgehog signalling, but thus far, the experimental results have been conflicting. To enable a variety of biochemical and structural experiments, we developed a method for expressing PTCHD1 in Spodoptera frugiperda cells, solubilising it in glycol-diosgenin, and purifying it to homogeneity. In vitro and in silico experiments show that PTCHD1 function is not interchangeable with Patched 1 (PTCH1) in canonical Hedgehog signalling, since it does not repress Smoothened in Ptch1-/- mouse embryonic fibroblasts and does not bind Sonic Hedgehog. However, we found that PTCHD1 binds cholesterol similarly to PTCH1. Furthermore, we identified 13 PTCHD1-specific protein interactors through co-immunoprecipitation and demonstrated a link to cell stress responses and RNA stress granule formation. Thus, our results support the notion that despite structural similarities to other Patched family proteins, PTCHD1 may have a distinct cellular function.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Hedgehog / Fibroblastos Limite: Animals Idioma: En Revista: Int J Mol Sci Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Finlândia País de publicação: Suíça

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Hedgehog / Fibroblastos Limite: Animals Idioma: En Revista: Int J Mol Sci Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Finlândia País de publicação: Suíça