Human receptor Smoothened, a mediator of Hedgehog signalling, expressed in its native conformation in yeast.
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.
De Rivoyre
Matthieu
M
Laboratoire de Physiologie Cellulaire et Moléculaire, UMR-CNRS 6548, Université de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2, France.
Bonino
Frédéric
F
Ruel
Laurent
L
Bidet
Michel
M
Thérond
Pascal
P
Mus-Veteau
Isabelle
I
D002846:Chromatography, Affinity; D015870:Gene Expression; D053823:Hedgehog Proteins; D006801:Humans; D010843:Pichia / Q000235:genetics*; D011487:Protein Conformation; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D015398:Signal Transduction; D000072086:Smoothened Receptor; D012995:Solubility; D013050:Spectrometry, Fluorescence; D015534:Trans-Activators / Q000378:metabolism*
D053823:Hedgehog Proteins; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; C491023:SMO protein, human; D000072086:Smoothened Receptor; D015534:Trans-Activators