1. Human receptor Smoothened, a mediator of Hedgehog signalling, expressed in its native conformation in yeast
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Frederic Bonino, Laurent Ruel, Pascal P. Thérond, Matthieu De Rivoyre, Isabelle Mus-Veteau, Michel Bidet, Physiologie cellulaire et moléculaire des systèmes intégrés (PCMSI), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Institut de signalisation, biologie du développement et cancer (ISBDC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)
- Subjects
MESH: Signal Transduction ,Protein Conformation ,Gene Expression ,MESH: Receptors, G-Protein-Coupled ,MESH: Pichia ,Biochemistry ,Chromatography, Affinity ,Pichia ,Receptors, G-Protein-Coupled ,MESH: Recombinant Proteins ,MESH: Protein Conformation ,0302 clinical medicine ,Structural Biology ,[SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB] ,MESH: Arthritis, Psoriatic ,Receptor ,MESH: Superoxide Dismutase ,Purification ,Smoothened ,0303 health sciences ,MESH: Gene Expression Regulation, Enzymologic ,MESH: RNA Stability ,MESH: Chromatography, Affinity ,Smoothened Receptor ,MESH: Saccharomyces cerevisiae ,Recombinant Proteins ,Signal transduction ,MESH: Spectrometry, Fluorescence ,Signal Transduction ,MESH: Cells, Cultured ,Patched ,MESH: Gene Expression ,MESH: Trans-Activators ,Saccharomyces cerevisiae ,Biophysics ,Biology ,Pichia pastoris ,03 medical and health sciences ,Mediator ,Genetics ,Humans ,Hedgehog Proteins ,Molecular Biology ,Hedgehog ,MESH: RNA, Messenger ,030304 developmental biology ,MESH: Tretinoin ,MESH: Humans ,MESH: Time Factors ,MESH: Hedgehog Proteins ,Cell Biology ,biology.organism_classification ,MESH: Male ,Yeast ,MESH: Solubility ,Spectrometry, Fluorescence ,Solubility ,MESH: Fibroblasts ,Membrane protein ,Hedgehog signalling ,Trans-Activators ,Heterologous expression ,MESH: Female ,030217 neurology & neurosurgery - Abstract
International audience; Though the role of Hedgehog (Hh) signalling in patterning and differentiation during development is well established, the underlying signal transduction mechanisms remain obscure. This is the first report on the overexpression of the human Hh signalling receptor Smoothened (hSmo) in Saccharomyces cerevisiae and Pichia pastoris. We show that hSmo is expressed in both types of yeast in its native conformational state. The first purification presented here will allow the characterisation of hSmo expressed in yeast, and the scale-up of hSmo production enabling structural studies to develop new therapeutic approaches against tumors and neurodegenerative diseases induced by Hh signalling dysfunction.
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