title: Heterologous expression of piglet odorant-binding protein in Pichia pastoris: a comparative structural and functional characterization with native forms.
abstract: This study targets to express the piglet odorant-binding protein (plOBP) and compare the engineered product to the corresponding native protein forms, i.e. plOBP and adult porcine OBP (pOBP). Using the natural signal peptide from the cDNA sequence, up to 40 mg l(-1) of secreted recombinant piglet OBP (rOBP) has been produced in a minimal culture medium. No significant difference in molecular mass between rOBP and native plOBP could be observed by mass spectrometry following or not trypsin digestion. rOBP and pOBP shared similar immunoreactivity towards polyclonal anti-pOBP antibodies, suggesting a proper processing and folding of the recombinant product. Both plOBP and rOBP displayed comparable binding properties towards fatty acids present in the putative maternal pheromone and a steroid, component of the boar sex pheromone. Furthermore, the rOBP product was found to bind to an olfactory receptor, for which pOBP binding was previously characterized. Taken together, these findings suggest that rOBP, produced in Pichia pastoris, exhibits structural and functional properties comparable to those of the native lipocalins from both young or adult animal.
authors.affiliation: Phérosynthèse SA, Le Rieu Neuf, F-84490 Saint-Saturnin-Les-Apt, France. gaelle.guiraudie-capraz@up.univ-mrs.fr
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Guiraudie-Capraz
authors.forename: Olivier
authors.initials: O
authors.lastname: Clot-Faybesse
authors.forename: Patrick
authors.initials: P
authors.lastname: Pageat
authors.forename: Christian
authors.initials: C
authors.lastname: Malosse
authors.forename: Anne-Hélène
authors.initials: AH
authors.lastname: Cain
authors.forename: Catherine
authors.initials: C
authors.lastname: Ronin
authors.forename: Patricia
authors.initials: P
authors.lastname: Nagnan-Le Meillour
mesh_terms: D000818:Animals; D010843:Pichia / Q000235:genetics*; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D013552:Swine
chemical_list: D018035:Receptors, Odorant; D011994:Recombinant Proteins; C045648:odorant-binding protein