Identification of a third rat odorant-binding protein (OBP3).
From a rat olfactory epithelium cDNA library clones encoding a lipocalin were isolated with sequence identity to the previously described salivary-specific alpha-2u globulin and the N-terminal region of mouse odorant-binding proteins OBP-III and OBP-IV. In situ hybridization showed strong expression in nasal glands displaying a pattern equivalent to rat OBP1. Heterologously expressed protein was evaluated for its binding properties using spectroscopic approaches. The recombinant protein interacted with two fluorescent probes, 1-aminoanthracene (1-AMA) and 1,1'-bis(4-anilino-5-naphthalene)-sulfonic acid. 1-AMA binding was competed by several odorants with high affinity. The thermodynamic parameters of the protein-odorant interaction were determined using isothermal titration calorimetry. Due to its nasal expression and odorant-binding characteristics this protein was designated OBP3.
Löbel
D
D
Institute of Physiology, University of Hohenheim, Stuttgart, Germany.
Strotmann
J
J
Jacob
M
M
Breer
H
H
D000817:Anilino Naphthalenesulfonates / Q000494:pharmacology; D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D002151:Calorimetry; D018076:DNA, Complementary / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D015723:Gene Library; D017403:In Situ Hybridization; D020128:Inhibitory Concentration 50; D008024:Ligands; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D018035:Receptors, Odorant / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013816:Thermodynamics
D000817:Anilino Naphthalenesulfonates; D000873:Anthracenes; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D008024:Ligands; D018035:Receptors, Odorant; D011994:Recombinant Proteins; C045648:odorant-binding protein; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); C049062:1-aminoanthracene