| Title: | Optimization of crystals from nanodrops: crystallization and preliminary crystallographic study of a pheromone-binding protein from the honeybee Apis mellifera L |
| Author(s): | Lartigue A; Gruez A; Briand L; Pernollet JC; Spinelli S; Tegoni M; Cambillau C; |
| Address: | "Architecture et Fonction des Macromolecules Biologiques, UMR 6098 CNRS and Universites Aix-Marseille I and 2, 31 Chemin Joseph Aiguier, 13402 Marseille CEDEX 20, France" |
| Journal Title: | Acta Crystallogr D Biol Crystallogr |
| DOI: | 10.1107/s090744490300413x |
| ISSN/ISBN: | 0907-4449 (Print) 0907-4449 (Linking) |
| Abstract: | "Pheromone-binding proteins (PBPs) are small helical proteins ( approximately 13-17 kDa) present in various sensory organs from moths and other insect species. They are involved in the transport of pheromones from the sensillar lymph to the olfactory receptors. Here, crystals of a PBP (Amel-ASP1) originating from honeybee (Apis mellifera L.) antennae and expressed as recombinant protein using the yeast Pichia pastoris are reported. Crystals of Amel-ASP1 have been obtained by the sitting-drop vapour-diffusion method using a nanodrop-dispensing robot under the following conditions: 200 nl of 40 mg ml(-1) protein solution in 10 mM Tris, 25 mM NaCl pH 8.0 was mixed with 100 nl of well solution containing 0.15 M sodium citrate, 1.5 M ammonium sulfate pH 5.5. The protein crystallizes in space group C222(1), with unit-cell parameters a = 74.8, b = 85.8, c = 50.2 A. With one molecule in the asymmetric unit, V(M) is 3.05 A(3) Da(-1) and the solvent content is 60%. A complete data set has been collected at 1.6 A resolution on beamline ID14-2 (ESRF, Grenoble). The nanodrop crystallization technique used with a novel optimization procedure made it possible to consume small amounts of protein and to obtain a unique crystal per nanodrop, suitable directly for data collection in-house or at a synchrotron-radiation source" |
| Keywords: | "Amino Acid Sequence Ammonium Sulfate/chemistry Animals Bees/*chemistry Carrier Proteins/*chemistry/genetics Crystallization/methods Crystallography, X-Ray Hydrogen-Ion Concentration *Insect Proteins Molecular Sequence Data Pichia/metabolism Recombinant Pr;" |
| Notes: | "MedlineLartigue, Audrey Gruez, Arnaud Briand, Loic Pernollet, Jean-Claude Spinelli, Silvia Tegoni, Mariella Cambillau, Christian eng Research Support, Non-U.S. Gov't 2003/06/05 Acta Crystallogr D Biol Crystallogr. 2003 May; 59(Pt 5):919-21. doi: 10.1107/s090744490300413x. Epub 2003 Apr 25" |
