title: Bacterial expression and conformational analysis of a chemosensory protein from Schistocerca gregaria.
abstract: Chemosensory proteins (CSPs) are a class of small, soluble proteins present at high concentrations in chemosensory organs of different insect species. Several pieces of evidence suggest their involvement in carrying chemical messages from the environment to chemosensory receptors. However, a structural description of the mechanism of delivery has not been reported. In order to provide the first detailed conformational characterization of these molecules, we cloned a specific isoform (CSP-sg4) from Schistocerca gregaria and expressed it in Escherichia coli. The product was obtained with yields of more than 20 mg per L of culture, all in its soluble form. The recombinant protein was identical to the native one with respect to pairing of the disulfide bridges, aggregative state and secondary structure elements. Structural investigations revealed a significantly stable polypeptide with respect to variations in temperature and acidity. CD analysis, preliminary NMR data and secondary structure prediction pointed to a correctly folded structure where helical regions and loops are alternated in a similar fashion as that observed for other classes of odorant- and pheromone-binding proteins presenting no sequence similarity to CSPs.
authors.affiliation: Dipartimento di Chimica, Universita' Federico II, Napoli, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Picone
authors.forename: O
authors.initials: O
authors.lastname: Crescenzi
authors.forename: S
authors.initials: S
authors.lastname: Angeli
authors.forename: S
authors.initials: S
authors.lastname: Marchese
authors.forename: A
authors.initials: A
authors.lastname: Brandazza
authors.forename: L
authors.initials: L
authors.lastname: Ferrara
authors.forename: P
authors.initials: P
authors.lastname: Pelosi
authors.forename: A
authors.initials: A
authors.lastname: Scaloni
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D006110:Grasshoppers / Q000737:chemistry*; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: C435683:CSP-sg4 protein, Schistocerca gregaria; D019476:Insect Proteins; D011994:Recombinant Proteins