Optimization of crystals from nanodrops: crystallization and preliminary crystallographic study of a pheromone-binding protein from the honeybee Apis mellifera L

Acta Crystallographica. Section D, Biological Crystallography
Audrey LartigueChristian Cambillau

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, suit...Continue Reading

Citations

Apr 4, 2006·Die Naturwissenschaften·Nicolette S Honson, Erika Plettner
Jan 24, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Julien StathopulosPhilippe Leone
May 30, 2008·Journal of Molecular Biology·Marion E PesentiChristian Cambillau
Dec 3, 2008·Journal of Bacteriology·Marina SiponenValérie Campanacci
Jul 27, 2018·Nature Communications·Alexander P HynesSylvain Moineau

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