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):

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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Dataset's files

Source:

			Action	DRS
	20180424_OBPn_LIQ_5_RB2_01_12840.d.rar	Other
	20180424_OBPn_LIQ_5_RB2_01_12840_DA4.3.110_1.mgf	Mgf
	20180424_OBPn_LIQ_5_RB2_01_12840_DA4.3.110_1.pride.mgf.gz	Mgf
	EF100X_RA1_01_16497.d.rar	Other
	OBP 100X CID.pride.mgf.gz	Mgf

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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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