Taxonomic distribution of proteins containing PhBP domains

The tree below includes only several representative species and genera. The complete taxonomic breakdown of all proteins containing PhBP domains can be accessed here. Click the counts or percentage values to display the corresponding proteins.

Protein structure encodes the ligand binding specificity in pheromonebinding proteins.

Du G, Prestwich GD
Biochemistry. 1995; 34: 8726-32

The ligand specificities and binding affinities of three recombinantpheromone binding proteins (PBPs) of two saturniid moths (genus Antheraea)were determined by using a novel binding assay in conjunction with twotritium-labeled constituents of the pheromone blend,[3H]-6E,11Z-hexadecadienyl acetate and [3H]-4E,9Z-tetradecadienyl acetate.The new binding assay, in which nonspecific adsorption to a plastic vesselis suppressed by presaturation of the surface with a 1-alkanol, allowsmeasurement of dissociation constants (KD) for lipophilic ligands fortheir carrier proteins. The three PBPs showed KD values for[3H]-6E,11Z-16:Ac and [3H]-4E,9Z-14:Ac between 0.6 and 30 microM, asdetermined by Scatchard analysis. Importantly, two PBPs (Aper-1 andAper-2) from one species showed opposite binding specificities for thesetwo ligands. Aper-1, like Apol-3, showed 15-fold higher affinity for6E,11Z-16:Ac than for 4E,9Z-14:Ac, while Aper-2 showed a 3.5-foldpreference for binding the shorter chain compound. In addition, for theApol-3 PBP, displacement of [3H]-6E,11Z-16:Ac binding by other pheromonecomponents or analogs showed a clear trend in relative binding affinity:6E,11Z-16:Ac > 4E,9Z-14:Ac > 6E,11Z-16:Al approximately 16:Ac >6E,11Z-16:OH > 4E,9Z-14:OH. These data clearly demonstrate a > 1000-foldrange of binding affinities among these very similar structures andunambiguously demonstrate the specificity of the PBP-pheromoneinteraction. Moreover, this assay offers the potential for determiningligand specificities for odorant binding proteins and other proteins inthe vertebrate lipocalin superfamily.

Bacterial expression and photoaffinity labeling of a pheromone bindingprotein.

Prestwich GD
Protein Sci. 1993; 2: 420-8

The first high-level production of a binding-active odorant bindingprotein is described. The expression cassette polymerase chain reactionwas used to generate a DNA fragment encoding the pheromone binding protein(PBP) of the male moth Antheraea polyphemus. Transformation of Escherichiacoli cells with a vector containing this construct generated clones which,when induced with isopropyl beta-D-thiogalactopyranoside, produced the14-kDa PBP in both the soluble fraction and in inclusion bodies.Purification of the soluble recombinant PBP by preparative isoelectricfocusing and gel filtration gave > 95% homogeneous protein, which wasimmunoreactive with an anti-PBP antiserum and exhibited specific,pheromone-displaceable covalent modification by the photoaffinity label[3H]6E,11Z-hexadecadienyl diazoacetate. Recombinant PBP wasindistinguishable from the insect-derived PBP, as determined by bothnative and denaturing gel electrophoresis, immunoreactivity, andphotoaffinity labeling properties. Moreover, the insoluble inclusion bodyprotein could be solubilized, refolded, and purified by the sameprocedures to give a recombinant PBP indistinguishable from the solublePBP. Proton NMR spectra of the soluble and refolded protein providefurther evidence that they possess the same folded structure.

Molecular cloning of an insect pheromone-binding protein.

Raming K, Krieger J, Breer H
FEBS Lett. 1989; 256: 215-8

Clones coding for the pheromone binding protein precursor have beenselected from a cDNA library derived from antennae of the male moth,Antheraea polyphemus. The deduced protein sequence consists of a signalpeptide of 20 amino acid residues and a mature binding protein of 142amino acid residues. RNA blot hybridization indicated that the mRNA isselectively expressed in the antennae of the male moth.