Functional studies of membrane-bound and purified human Hedgehog receptor Patched expressed in yeast
| Autor: | Matthieu De Rivoyre, Olivier Joubert, Patrick Mollat, Bernard Pucci, Rony Nehmé, Damien Fleury, Isabelle Mus-Veteau, Michel Bidet, Martial Ruat, Ange Polidori |
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| Přispěvatelé: | Biologie et physiopathologie des systèmes intégrés (LBPSI), Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Galapagos SASU, Laboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels (LCBOSMV), Avignon Université (AU), Institut de Neurobiologie Alfred Fessard (INAF), Centre National de la Recherche Scientifique (CNRS), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM) |
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Patched
Patched Receptors endocrine system diseases Protein Conformation Biophysics Receptors Cell Surface Saccharomyces cerevisiae Membrane receptor MESH: Receptors G-Protein-Coupled Biology Biochemistry Receptors G-Protein-Coupled 03 medical and health sciences MESH: Protein Conformation Cell surface receptor Humans Hedgehog Proteins Sonic hedgehog Receptor MESH: Peptide Fragments 030304 developmental biology MESH: Receptors Cell Surface 0303 health sciences MESH: Humans 030302 biochemistry & molecular biology Cell Membrane Cell Biology MESH: Hedgehog Proteins Surface Plasmon Resonance Smoothened Receptor MESH: Saccharomyces cerevisiae Hedgehog signaling pathway Peptide Fragments MESH: Surface Plasmon Resonance Patched-1 Receptor Sonic Hedgehog biology.protein [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] Heterologous expression Smoothened Fluorinated surfactants MESH: Cell Membrane |
| Zdroj: | Biochimica et Biophysica Acta-Molecular Cell Research Biochimica et Biophysica Acta-Molecular Cell Research, Elsevier, 2009, 1788 (9), pp.1813-21. ⟨10.1016/j.bbamem.2009.05.009⟩ |
| ISSN: | 0167-4889 |
| Popis: | International audience; The Sonic Hedgehog (Shh) signalling pathway plays an important role both in embryonic development and in adult stem cell function. Inappropriate regulation of this pathway is often due to dysfunction between two membrane receptors Patched (Ptc) and Smoothened (Smo), which lead to birth defects, cancer or neurodegenerative diseases. However, little is known about Ptc, the receptor of the Shh protein, and the way Ptc regulates Smo, the receptor responsible for the transduction of the signal. To develop structure-function studies of these receptors, we expressed human Ptc (hPtc) in the yeast Saccharomyces cerevisiae. We demonstrated that hPtc expressed in a yeast membrane fraction is able to interact with its purified ligand Shh, indicating that hPtc is produced in yeast in its native conformational state. Using Surface Plasmon Resonance technology, we showed that fluorinated surfactants preserve the ability of hPtc to interact with its ligand after purification. This is the first report on the heterologous expression and the purification of a native and stable conformation of the human receptor Ptc. This work will allow the scale-up of hPtc production enabling its biochemical characterization, allowing the development of new therapeutic approaches against diseases induced by Shh signalling dysfunction. |
| Databáze: | OpenAIRE |
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