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Eur J Biochem

Title:		Characterization of a chemosensory protein (ASP3c) from honeybee (Apis mellifera L.) as a brood pheromone carrier
Author(s):		Briand L; Swasdipan N; Nespoulous C; Bezirard V; Blon F; Huet JC; Ebert P; Penollet JC;
Address:		"Biochimie et Structure des Proteines, Unite de recherches INRA 477, Jouy-en-Josas Cedex, France"
Journal Title:		Eur J Biochem
Year:		2002
Volume:		269
Issue:		18
Page Number:		4586 - 4596
DOI:		10.1046/j.1432-1033.2002.03156.x
ISSN/ISBN:		0014-2956 (Print) 0014-2956 (Linking)
Abstract:		"Chemosensory proteins (CSPs) are ubiquitous soluble small proteins isolated from sensory organs of a wide range of insect species, which are believed to be involved in chemical communication. We report the cloning of a honeybee CSP gene called ASP3c, as well as the structural and functional characterization of the encoded protein. The protein was heterologously secreted by the yeast Pichia pastoris using the native signal peptide. ASP3c disulfide bonds were assigned after trypsinolysis followed by chromatography and mass spectrometry combined with microsequencing. The pairing (Cys(I)-Cys(II), Cys(III)-Cys(IV)) was found to be identical to that of Schistocerca gregaria CSPs, suggesting that this pattern occurs commonly throughout the insect CSPs. CD measurements revealed that ASP3c mainly consists of alpha-helices, like other insect CSPs. Gel filtration analysis showed that ASP3c is monomeric at neutral pH. Using ASA, a fluorescent fatty acid anthroyloxy analogue as a probe, ASP3c was shown to bind specifically to large fatty acids and ester derivatives, which are brood pheromone components, in the micromolar range. It was unable to bind tested general odorants and other tested pheromones (sexual and nonsexual). This is the first report on a natural pheromonal ligand bound by a recombinant CSP with a measured affinity constant"
Keywords:		"Amino Acid Sequence Animals Base Sequence Bees/genetics/*metabolism Circular Dichroism DNA, Complementary Disulfides/metabolism Fluorescence Fluorescent Dyes Insect Proteins/*metabolism Ligands Molecular Sequence Data Pheromones/*metabolism Sequence Align;"
Notes:		"MedlineBriand, Loic Swasdipan, Nicharat Nespoulous, Claude Bezirard, Valerie Blon, Florence Huet, Jean-Claude Ebert, Paul Penollet, Jean-Claude eng England 2002/09/17 Eur J Biochem. 2002 Sep; 269(18):4586-96. doi: 10.1046/j.1432-1033.2002.03156.x"