Title: | Sulfur single-wavelength anomalous diffraction crystal structure of a pheromone-binding protein from the honeybee Apis mellifera L |
Author(s): | Lartigue A; Gruez A; Briand L; Blon F; Bezirard V; Walsh M; Pernollet JC; Tegoni M; Cambillau C; |
Address: | "Architecture et Fonction des Macromolecules Biologiques, Unite Mixte de Recherche 6098 CNRS and Universites Aix-Marseille I and II, 13402 Marseille Cedex 20, France" |
ISSN/ISBN: | 0021-9258 (Print) 0021-9258 (Linking) |
Abstract: | "Pheromone binding proteins (PBPs) are small helical proteins ( approximately 13-17 kDa) present in several sensory organs from moth and other insect species. They are involved in the transport of pheromones from the sensillar lymph to the olfactory receptors. We report here the crystal structure of a PBP (Amel-ASP1) originating from the honey-bee (Apis mellifera) antennae and expressed as recombinant protein in the yeast Pichia pastoris. Crystals of Amel-ASP1 were obtained at pH 5.5 using the nano-drops technique of crystallization with a novel optimization procedure, and the structure was solved initially with the single-wavelength anomalous diffraction technique using sulfur anomalous dispersion. The structure of Amel-ASP1 has been refined at 1.6-A resolution. Its fold is roughly similar to that of other PBP/odorant binding proteins, presenting six helices and three disulfide bridges. Contrary to the PBPs from Bombyx mori (Sandler, B. H., Nikonova, L., Leal, W. S., and Clardy, J. (2000) Chem. Biol. 7, 143-151) and Leucophea maderae (Lartigue, A., Gruez, A., Spinelli, S., Riviere, S., Brossut, R., Tegoni, M., and Cambillau, C. (2003) J. Biol. Chem. 278, 30213-30218), the extended C terminus folds into the protein and forms a wall of the internal hydrophobic cavity. Its backbone groups establish two hydrogen bonds with a serendipitous ligand, n-butyl-benzene-sulfonamide, an additive used in plastics. This mode of binding might, however, mimic that used by one of the pheromonal blend components and illustrates the binding versatility of PBPs" |
Keywords: | "Animals Bees/chemistry/genetics Carrier Proteins/*chemistry/genetics Crystallization Crystallography, X-Ray Insect Proteins/*chemistry/genetics Models, Molecular Protein Conformation Recombinant Proteins/chemistry/genetics Static Electricity Sulfur/chemis;" |
Notes: | "MedlineLartigue, Audrey Gruez, Arnaud Briand, Loic Blon, Florence Bezirard, Valerie Walsh, Martin Pernollet, Jean-Claude Tegoni, Mariella Cambillau, Christian eng Research Support, Non-U.S. Gov't 2003/11/05 J Biol Chem. 2004 Feb 6; 279(6):4459-64. doi: 10.1074/jbc.M311212200. Epub 2003 Oct 31" |