Dataset Information

Binding specificity of Native Odorant-Binding Protein isoforms is driven by phosphorylation and O-N-acetylglucosamiylation in the pig (sus scrofa)

ABSTRACT: In pig, phosphorylation drives binding specificity of recombinant OBP (stricto sensu, referred as OBP in this text) isoforms (Brimau et al., 2010) that are not O-GlcNAcylated, contrary to native OBP, purified from nasal tissue (Nagnan-Le Meillour et al., 2014). In order to precise the role of both PTM in OBP binding abilities to pheromone components in pig, we purified OBP isoforms by HPLC and performed structure-function relationship study. We have identified phosphorylation and O-GlcNAcylation on 4 OBP isoforms by immunodetection with specific antibodies, with careful controls. The binding properties of the 4 isoforms were monitored by fluorescence spectroscopy in exactly the same conditions previously used for recombinant OBP isoforms (Brimau et al., 2010). The PTM sites were mapped by CID-nano-LC-MS/MS and BEMAD (for phosphorylation) to link the binding affinities to phosphorylation and O-GlcNAcylation patterns of native OBP isoforms. Comparison of binding affinities between recombinant (only phosphorylated) and native (also O-GlcNAcylated) isoforms indicates that O-GlcNAcylation increases the binding specificity to pheromone components. This is the first time that such PTM sites are localized in OBP by high-resolution mass spectrometry, and that a demonstration of their involvement in odorant molecules discrimination by OBP isoforms is given

INSTRUMENT(S): maXis

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Respiratory Mucus

SUBMITTER: Alexandre Joly

LAB HEAD: Patricia Nagnan-Le Meillour

PROVIDER: PXD011371 | Pride | 2019-02-15

REPOSITORIES: Pride

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Publications

Binding Specificity of Native Odorant-Binding Protein Isoforms Is Driven by Phosphorylation and <i>O</i>-N-Acetylglucosaminylation in the Pig <i>Sus scrofa</i>.

Nagnan-Le Meillour Patricia P Joly Alexandre A Le Danvic Chrystelle C Marie Arul A Zirah Séverine S Cornard Jean-Paul JP

Frontiers in endocrinology 20190125

Odorant-binding proteins (OBP) are secreted in the nasal mucus at the vicinity of olfactory receptors (ORs). They act, at least, as an interface between hydrophobic and volatile odorant molecules and the hydrophilic medium bathing the ORs. They have also been hypothesized to be part of the molecular coding of odors and pheromones, by forming specific complexes with odorant molecules that could ultimately stimulate ORs to trigger the olfactory transduction cascade. In a previous study, we have ev ...[more]

PMID: 30740091

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Project description:Background: Small ungulates (sheep and goat) display a seasonal breeding, characterised by successive periods of sexual activity (SA) and sexual rest (SR). In sexual activity period the ovarian cycle of females is active and ready for reproduction (oestrus) whereas in sexual rest no ovulation and reproduction are possible (deep anoestrus). Odours emitted by a sexually active male can reactivate the ovarian cycle of anoestrus females. The plasticity of the olfactory system under these hormonal changes has never been explored at the peripheral level of odours reception. As it was shown in pig that the olfactory secretome (proteins secreted in the nasal mucus) could be modified under hormonal control, we monitored its composition in females of both species along several seasons, thanks to a non-invasive sampling of olfactory mucus. Results: In both species the olfactory secretome is composed of isoforms of OBP-like proteins, generated by post-translational modifications, phosphorylation, N-glycosylation and O-GlcNAcylation. Important changes were observed in the olfactory secretome between the sexual rest and the sexual activity periods, characterised in ewe by the specific expression of SAL-like proteins and the emergence of OBPs O-GlcNAcylation. In Goat, the differences between SA and SR did not come from new proteins expression, but from different post-translational modifications, the main difference between the SA and SR secretome being the number of isoforms of each protein. Conclusion: Despite common behaviour, seasonal breeding, and genetic resources, the two species seem to adapt their sensory equipment in SA by different modalities: the variation of olfactory secretome in ewe could correspond to a specialization to detect male odours only in SA, whereas in goat the stability of the olfactory secretome could indicate a constant capacity of odours detection suggesting that the hallmark of SA in goat could be the emission of specific odours by the sexually active male.

Dataset Information

Binding specificity of Native Odorant-Binding Protein isoforms is driven by phosphorylation and O-N-acetylglucosamiylation in the pig (sus scrofa)

Publications

Binding Specificity of Native Odorant-Binding Protein Isoforms Is Driven by Phosphorylation and <i>O</i>-N-Acetylglucosaminylation in the Pig <i>Sus scrofa</i>.

Similar Datasets

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