title: Properties of mouse vomeronasal receptor and assessment of its role in pheromone signalling.
abstract: Vomeronasal type 2 receptor (V2Rx) from Swiss mouse (Mus musculus (L.)) was analyzed by high-resolution ion-exchange chromatography, reversed-phase high-performance liquid chromatography (RP-HPLC), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), Ion Spray tandem mass spectrometry (MS/MS), 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and 1-aminoanthracene (1-AMA) fluorometric assay. Vomeronasal sensory neuronal cell bound proteins were resolved into major protein peaks. Several proteins were identified and subsequently purified as the V2Rx receptor on 10% SDS-PAGE with trace amounts of other protein bands. The molecular weight of the identified V2Rx was 109 kDa. MALDI-TOF and micro-sequencing experiments demonstrated that the identified V2Rx receptor shared considerable sequence similarity with vomeronasal receptor type 2 (NCBI Accession Number AB267725), which is a seven transmembrane peptide with 912 amino acid residues. The molecular characterization revealed that the N-terminus of the V2Rx receptor contained the 11GAEAAE16 domain involved in pheromone signalling. The biometric assay (octanamine-V2Rx binding) showed the identified V2Rx receptor and mouse sex pheromone to 2-octanamine (methyl heptyl) in a 1:1 ratio. Uptake of odourants determined in physiological condition showed enhanced V2Rx receptors as volatile hydrophobic pheromone receptors in the vomeronasal neuron of the Swiss mouse.
authors.affiliation: Proteomics & Molecular Cell Physiology Laboratory, Department of Zoology, Bharathiar University, Coimbatore-641 046, TN, India. sk_protein@buc.edu.in
authors.forename: Soundarapandian
authors.initials: S
authors.lastname: Kannan
authors.forename: Pandi
authors.initials: P
authors.lastname: Mareeswaran
authors.forename: Muthukalingan
authors.initials: M
authors.lastname: Krishnan
authors.forename: Shanmugam
authors.initials: S
authors.lastname: Achiraman
authors.forename: Neil
authors.initials: N
authors.lastname: Spencer
authors.forename: Balakrishnan F
authors.initials: BF
authors.lastname: Nytingel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000873:Anthracenes / Q000737:chemistry / Q000378:metabolism; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D043563:Receptors, Pheromone / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D019147:Vomeronasal Organ / Q000737:chemistry*
chemical_list: D000873:Anthracenes; D043563:Receptors, Pheromone; C049062:1-aminoanthracene