Natural Ligands of Porcine Olfactory Binding Proteins

Danvic, Chrystelle Le; Guiraudie-Capraz, Gaëlle; Abderrahmani, Djamel; Zanetta, Jean‐Pierre; Meillour, Patricia Nagnan‐Le

doi:10.1007/s10886-009-9645-1

Cited by 23 publications

(23 citation statements)

References 28 publications

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“…Recombinant OBPs from P. pastoris, isoforms or total protein, did not bind testosterone. This result was expected from the in-gel binding assay with native OBP that did not bind testosterone (Le Danvic et al 2009), which is the natural ligand of VEG (Von Ebner's Gland protein), another protein of the nasal mucus. Whatever the ligand, OBP-Pichia-iso2 displayed the same behavior as total OBP-Pichia; that could be explained by the fact that it is the closest to total OBPPichia, in terms of phosphorylation.…”

Section: Ligand Binding Monitored By Fluorescence Spectroscopymentioning

confidence: 57%

“…In addition, we demonstrated recently that a dynamic mechanism involving PTM could be responsible for OBP diversity and specificity (Le Danvic et al 2009;Nagnan-Le Meillour et al 2009b). Indeed, porcine OBP can be posttranslationally modified by phosphorylation, and we suggested that this PTM could generate a diversity of OBP isoforms with specific binding properties (Nagnan-Le Meillour et al 2009b).…”

Section: Introductionmentioning

confidence: 71%

“…For adult females, androstenone is a sex pheromone, but for piglets, the perception of androstenone reduces agonistic behaviors and is considered as a submissive pheromone (McGlone 1985;McGlone and Morrow 1988). Testosterone, the natural ligand of VEG (a pheromone binding protein) (Le Danvic et al 2009), is never bound by any of the OBP isoforms. Conversely, both OBP isoforms bind a closely related steroid, androstenone, differing from testosterone by a single alcohol.…”

Section: Ligand Binding Monitored By Fluorescence Spectroscopymentioning

confidence: 99%

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Binding Specificity of Recombinant Odorant-Binding Protein Isoforms is Driven by Phosphorylation

et al. 2010

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Native porcine odorant-binding protein (OBP) bears eleven sites of phosphorylation, which are not always occupied in the molecular population, suggesting that different isoforms could co-exist in animal tissues. As phosphorylation is a dynamic process resulting in temporary conformational changes that regulate the function of target proteins, we investigated the possibility that OBP isoforms could display different binding affinities to biologically relevant ligands. The availability of recombinant proteins is of particular interest for the study of protein/ligand structure-function relationships, but prokaryotic expression systems do not perform eukaryotic post-translational modifications. To investigate the role of phosphorylation in the binding capacities of OBP isoforms, we produced recombinant porcine OBP in two eukaryotic systems, the yeast, Pichia pastoris, and the mammalian CHO cell line. Isoforms were separated by anion exchange HPLC, and their phosphorylation sites were mapped by MALDI-TOF mass spectrometry and compared to those of the native protein. Binding experiments with ligands of biological relevance in the pig, Sus scrofa, were performed by fluorescence spectroscopy on two isoforms of recombinant OBP expressed in the yeast. The two isoforms, differing only by their phosphorylation pattern, displayed different binding properties, suggesting that binding specificity is driven by phosphorylation.

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Section: Ligand Binding Monitored By Fluorescence Spectroscopymentioning

confidence: 57%

Section: Introductionmentioning

confidence: 71%

Section: Ligand Binding Monitored By Fluorescence Spectroscopymentioning

confidence: 99%

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Binding Specificity of Recombinant Odorant-Binding Protein Isoforms is Driven by Phosphorylation

et al. 2010

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“…Solution-based affinity binding is commonly characterized using methods such as UV-absorption [8], differential and titration calorimetry [9], matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) [10] and electrophoretic separation [11]. Surface-based affinity binding, which is widely used in affinity biosensors [12], can be investigated with methods such as protein arrays [13], immunoassays [14], and thermal-shift assays [15], which use fluorescent or radioactive labeling groups to signal binding events.…”

Section: Introductionmentioning

confidence: 99%

Microcantilever-based label-free characterization of temperature-dependent biomolecular affinity binding

Wang

Huang

Nguyen

et al. 2013

Sensors and Actuators B: Chemical

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This paper presents label-free characterization of temperature-dependent biomolecular affinity binding on solid surfaces using a microcantilever-based device. The device consists of a Parylene cantilever one side of which is coated with a gold film and functionalized with molecules as an affinity receptor to a target analyte. The cantilever is located in a poly(dimethylsiloxane) (PDMS) microfluidic chamber that is integrated with a transparent indium tin oxide (ITO) resistive temperature sensor on the underlying substrate. The ITO sensor allows for real-time measurements of the chamber temperature, as well as unobstructed optical access for reflection-based optical detection of the cantilever deflection. To test the temperature-dependent binding between the target and receptor, the temperature of the chamber is maintained at a constant setpoint, while a solution of unlabeled analyte molecules is continuously infused through the chamber. The measured cantilever deflection is used to determine the target-receptor binding characteristics. We demonstrate label-free characterization of temperature-dependent binding kinetics of the platelet-derived growth factor (PDGF) protein with an aptamer receptor. Affinity binding properties including the association and dissociation rate constants as well as equilibrium dissociation constant are obtained, and shown to exhibit significant dependencies on temperature.

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“…The calyx is a barrel structure composed of eight β‐sheets in alternating reverse orientation, with α‐helix linkages between some of the individual sheets (Grzyb et al, 2006). The binding of lipocalins to specific ligands plays a role in many biological functions, including olfaction (Lazer et al, 2002; Le Danvic et al, 2009), coloration (Cianci et al, 2002; Wade et al, 2009), immune response (Logdberg and Wester, 2000; Clifton et al, 2009), pheromone transport (Miyawaki et al, '95; Magert et al, '99; Chamero et al, 2007), prostaglandin synthesis (Nagata et al, '91; Fouchecourt et al, 2002) and allergen response (Virtanen et al, '99; Virtanen, 2001). Retinol binding protein (RBP), which is found in human, rat and other species, is the canonical member of the lipocalin family.…”

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confidence: 99%

Prawn lipocalin: characterization of a color shift induced by gene knockdown and ligand binding assay

Yang

Wang

et al. 2011

J. Exp. Zool.

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The lipocalin family of proteins functions in the transport of steroids, carotenoids, retinoids, and other small hydrophobic molecules. Recently, a lipocalin (MrLC) was isolated from the prawn Macrobrachium rosenbergii and its expression varied with the molting cycle. In this study, knockdown of the MrLC gene by RNA interference (RNAi) was performed and resulted in a shift in body color from blue to orangish red over the entire carapace. By immune-gold electron microscopy, MrLC was found to co-localize with the lipid droplets in subepidermal adiose tissue that were found to be decreased dramatically in MrLC knockdown prawns, in which a reduction in relative fat content was also quantified. Furthermore, MrLC was found to specifically bind astaxanthin and molt hormone (20-hydroxyecdysone) in both in vitro ligand binding assay and in vivo native ligand detection. These results suggested that MrLC plays roles in the regulation of coloration through its association with astaxanthin and may also be involved in the regulation of molting in crustacean.

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Natural Ligands of Porcine Olfactory Binding Proteins

Cited by 23 publications

References 28 publications

Binding Specificity of Recombinant Odorant-Binding Protein Isoforms is Driven by Phosphorylation

Binding Specificity of Recombinant Odorant-Binding Protein Isoforms is Driven by Phosphorylation

Microcantilever-based label-free characterization of temperature-dependent biomolecular affinity binding

Prawn lipocalin: characterization of a color shift induced by gene knockdown and ligand binding assay

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