Cloning, post-translational modifications, heterologous expression and ligand-binding of boar salivary lipocalin

LOEBEL, Dietrich; Scaloni, Andrea; Paolini, Sara; Fini, Carlo; Ferrara, L.; Breer, Heinz; Pelosi, Paolo

doi:10.1042/0264-6021:3500369

Cited by 35 publications

(32 citation statements)

References 34 publications

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“…1). In particular, AimelOBP3 is about 54% identical with pig SALs, which are responsible for carrying the sex pheromones androstenone and androstenol in the saliva of the boar as well as for detecting them in the nose (20)(21)(22). They belong to a subgroup including the rodent MUPs (18,31), the hamster aphrodisin (32), and rabbit seminal protein OBP3 (33), which are all involved in the release of specific pheromones (11).…”

Section: Resultsmentioning

confidence: 99%

“…The most compelling facts are their sites of production outside the olfactory mucosa in the vomeronasal organ and the nasal respiratory epithelium and the occurrence of the same or very similar OBPs in the nose and in biological glands and fluids releasing specific pheromones. The best examples of this fact are the major urinary proteins (MUPs) of rodents (18,19) and the salivary lipocalins (SALs) of the pig (20)(21)(22). In both cases, the same proteins are produced in the nose as well as in the liver (in rodents) or in the salivary glands (in the pig).…”

Section: Significancementioning

confidence: 99%

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Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles

Zhu

Arena

Spinelli

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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104

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The giant panda Ailuropoda melanoleuca belongs to the family of Ursidae; however, it is not carnivorous, feeding almost exclusively on bamboo. Being equipped with a typical carnivorous digestive apparatus, the giant panda cannot get enough energy for an active life and spends most of its time digesting food or sleeping. Feeding and mating are both regulated by odors and pheromones; therefore, a better knowledge of olfaction at the molecular level can help in designing strategies for the conservation of this species. In this context, we have identified the odorant-binding protein (OBP) repertoire of the giant panda and mapped the protein expression in nasal mucus and saliva through proteomics. Four OBPs have been identified in nasal mucus, while the other two were not detected in the samples examined. In particular, AimelOBP3 is similar to a subset of OBPs reported as pheromone carriers in the urine of rodents, saliva of the boar, and seminal fluid of the rabbit. We expressed this protein, mapped its binding specificity, and determined its crystal structure. Structural data guided the design and preparation of three protein mutants bearing single-amino acid replacements in the ligand-binding pocket, for which the corresponding binding affinity spectra were measured. We also expressed AimelOBP5, which is markedly different from AimelOBP3 and complementary in its binding spectrum. By comparing our binding data with the structures of bamboo volatiles and those of typical mammalian pheromones, we formulate hypotheses on which may be the most relevant semiochemicals for the giant panda.odorant-binding proteins | chemical communication | X-ray structure | proteomics | giant panda T he giant panda Ailuropoda melanoleuca is endemic of China and was formerly classified as an endangered species, now as a vulnerable species, but its population has remained rather stable, although very low, during the last centuries (1). Its phylogenetic classification has been a matter of debate for some time, but molecular genetic studies have recently shown that this species belongs to Ursidae, of which it represents an ancestral branch together with the spectacled bears, Tremarctos, and the sloth bear (1-3). The diets of these species are different from those of carnivorous bears: the giant panda is fully herbivorous, the spectacled bears are mainly herbivorous, and sloth bears feed on termites, fruits, and vegetables. The giant panda also shares with spectacled bears and sloth bears the absence of hibernation, an important characteristic that differentiates these species from other Ursidae (4).The obligate bamboo diet of the giant panda, which is not compatible with its carnivorous digestive system, is barely sufficient to provide the energy required for an active life, likely accounting for the slow movements and long periods of rest typical of this species (5). It has been also suggested that the reduced size of the brain, liver, and kidneys of the giant panda relative to other mammals could be a measure to further reduce the use of i...

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Section: Resultsmentioning

confidence: 99%

Section: Significancementioning

confidence: 99%

Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles

Zhu

Arena

Spinelli

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

104

View full text Add to dashboard Cite

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“…Fluorescence Binding Assay with AMA and Competition between AMA and 1-Octen 3-ol-The fluorescence properties of AMA when surrounded by a hydrophobic environment have already been used to probe binding in pOBP (20) and porcine salivary OBP (34). We have determined the binding affinity of AMA for ligand-depleted bOBP by titrating at 480 nm the increase in fluorescence signal of AMA upon complexation to the protein (Fig.…”

Section: Fig 1 Gas Chromatography-coupled Mass Spectroscopy Of the mentioning

confidence: 99%

The Insect Attractant 1-Octen-3-ol Is the Natural Ligand of Bovine Odorant-binding Protein

Ramoni

Vincent

Grolli

et al. 2001

Journal of Biological Chemistry

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“…Among liver-specific proteins, there were some housekeeping proteins, and there were several others already reported to be abundantly expressed in liver, including catalase, cytochrome P450, and ornithine carbamoyltransferase (Lin et al, 1997;Baranova et al, 2005;Koger and Jones, 1997). Likewise, among the salivary gland-specific proteins, we identified some predicted to function in the salivary gland or saliva, such as submaxillary apomucin and salivary lipocalin precursor (Eckhardt et al, 1997;Loebel et al, 2000). Although we did not identify a high number of organ-specific proteins, comparison of the proteomes of salivary gland, liver, and differentiated SGP may allow determination of the state of SGP differentiation.…”

Section: Discussionmentioning

confidence: 92%

Large-Scale Identification by Shotgun Proteomics of Proteins Expressed in Porcine Liver and Salivary Gland

et al. 2008

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Protein catalogs containing a large number of proteins expressed in a variety of organs can be powerful tools for stem-cell research, because this requires accurate knowledge about how cells differentiate. Salivary gland progenitor (SGP) cells are somatic stem cells isolated from the salivary gland that can differentiate into hepatic or pancreatic cell lineages. Their differentiation state has been assessed by the expression of major protein markers, but to use these cells in regenerative medicine, it will be necessary to establish additional means of quality assessment. We examined the use of shotgun proteomics for porcine salivary gland (a source of SGP cells) and liver (a destination of differentiated SGP cells) for determining the state of SGP cell differentiation. Protein complexes from each organ were digested into peptides and separated by two-dimensional liquid chromatography involving strong cation-exchange chromatography followed by reversed-phase liquid chromatography. The separated peptides were analyzed by on-line electrospray ionization tandem mass spectrometry using a quadrupole-time of flight mass spectrometer (ESI Q-TOF MS/ MS), and the spectra obtained were processed to search peptides against a mammalian database for protein identification. Using this method, we identified 117 proteins in porcine salivary gland and 154 proteins in porcine liver. Of these, 72 and 109 were specific to salivary gland and liver, respectively, and some of these were previously shown to be organ specific. The current study can be utilized in the future as a basis to study the pattern of differentiation in protein expression by stem cells.

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Cloning, post-translational modifications, heterologous expression and ligand-binding of boar salivary lipocalin

Cited by 35 publications

References 34 publications

Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles

Reverse chemical ecology: Olfactory proteins from the giant panda and their interactions with putative pheromones and bamboo volatiles

The Insect Attractant 1-Octen-3-ol Is the Natural Ligand of Bovine Odorant-binding Protein

Large-Scale Identification by Shotgun Proteomics of Proteins Expressed in Porcine Liver and Salivary Gland

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