Odorant and pheromone binding by aphrodisin, a hamster aphrodisiac protein

Briand, Loı̈c; Huet, Jean‐Claude; Pérez, Valérie; Lenoir, Guillaume; Nespoulous, Claude; Boucher, Yves; Trotier, Didier; Pernollet, Jean‐Claude

doi:10.1016/s0014-5793(00)01719-1

Cited by 58 publications

(42 citation statements)

References 44 publications

(53 reference statements)

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“…The molecular weight of the overexpressed Aphrodisin in the present study was ~17kDa, which is similar to earlier reports of molecular weight of natural aphrodisin of Syrian hamster (Singer et al, 1986;Magert et al, 1999;Briand et al, 2000). Peptide mass fingerprinting and matrix-assisted laser-desorption ionization-time-offlight (MALDI-TOF) mass spectra analysis of the trypsin digested ~17 kDa recombinant protein, and MS/MS analysis indicated that the overexpressed ~17 kDa protein was closely similar to aphrodisin of Syrian hamster (Figure-8 a-b).…”

Section: Tryptic Digestion and Mass-spectrometric Analysis Of The ~17supporting

confidence: 92%

“…Lipocalins display a remarkable range of different molecular recognition properties and are variable in sequence, but share a typical 3D structure (Flower et al, 1993;Flower, 1995 and1996). Among lipocalins, aphrodisin is highly homologous (40% sequence identity) to rat odorant binding proteins OBP-1 and IF, expressed and secreted by nasal glands of rat in both sexes, which are thought to carry volatile hydrophobic odorants towards olfactory receptors across the aqueous nasal mucus (Pevsner et al, 1988;Briand et al, 2000). Aphrodisin has also 39% identity with a male-specific lipocalin (MSP) expressed in submandibular glands of male hamster, which is secreted in saliva (Thavathiru et al, 1999;Srikantan et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Determination of Aphrodisin Gene of Syrian Hamster by Molecular Technique

Khan

2012

Bangl. J. Vet. Med.

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Aphrodisin is a secretory protein expressed in the reproductive organs and parotid salivary gland of the female hamster and acts as an aphrodisiac pheromone. RT-PCR and cloning was applied for the determination of aphrodisin gene from the vaginal tissues of the hamster. In the present study, total RNA was extracted from hamster uterus by Trizol method and apparently undegraded 28S, 18S and 5S species of ribosomal RNA was clearly visible. After performing RT-PCR a distinct band of ~475 bp was obtained and showed in electrophoresis gel analysis which was the partial cDNA of aphrodisin protein. After purification of PCR product and its sequencing using OE-F and OE-R primers confirmed the PCR product was the full-length cDNA of mature aphrodisin of 151 amino acids. The pET 21 plasmid purified from DH5-α cell and ligated with Aph-insert of ~475 bp. Subsequent sequencing confirmed error free ligation and presence of aphrodisin insert. No sequence discrepancy was noted with the published cDNA sequence of mature aphrodisin except for a single base mismatch (GGg for GGA), which did not result in any change in the coded amino acid (glycine). SDS-PAGE analysis showed a major protein greater than 17-kDa was observed in the protein profiles of post-lysis pellet of IPTG induced BL-21 cells but not in their lysates. This 17 kDa protein band should represent recombinant aphrodisin protein which is present in the post-lysis pellet since it is not solubilized by simple sonication. Peptide mass fingerprinting and matrix-assisted laser-desorption ionization-time-of-flight (MALDI-TOF) mass spectra analysis of the trypsin digested ~17 kDa recombinant protein, and mass spectrophotometrical (MS) analysis indicated that the overexpressed ~17 kDa protein was closely similar to aphrodisin of Syrian hamster. Key words: Syrian hamster, aphrodisin protein, overexpression INTRODUCTIONAphrodisin is a 17-kDa soluble glycosylated secretory protein expressed in vagina, uterus, Bartholin's gland of the female hamster and also in the female's parotid salivary glands (Magert et al., 1999), and in the bank vole (Stopkova et al., 2010). Aphrodisin acts as an aphrodisiac pheromone that is deposited upon the snout and tongue of the male by contact during normal chemosensory investigation of the female's vaginal discharge by the male hamster, and it is probably transported in the nasal mucus to chemoreceptors in the male's vomeronasal organ (Clancy et al., 1984).Aphrodisin consists of 151 amino acids with two disulfide bonds and a blocked N-terminus (pyroglutamic acid). Each N-glycosylation site (N41 and N69) is linked with only one N-acetylglucosamine residue (Singer et al., 1986). Aphrodisin belongs to lipocalin family, members of which are known to be carrier proteins and they transport low molecular weight hydrophobic molecules. Lipocalins display a remarkable range of different molecular recognition properties and are variable in sequence, but share a typical 3D structure (Flower et al., 1993;Flower, 1995 and1996). Among lipocalins, aphrodis...

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Section: Tryptic Digestion and Mass-spectrometric Analysis Of The ~17supporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Determination of Aphrodisin Gene of Syrian Hamster by Molecular Technique

Khan

2012

Bangl. J. Vet. Med.

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“…Nevertheless, once cleaved from the glutamine residue as shown in this work, the acids might subsequently associate non-covalently with apolipoprotein D, and this non-covalent association could be just one more mechanism of slow release of these volatiles from the axilla. A similar release mechanism is postulated for hamster aphrodisin and a putative hydrophobic pheromonal compound (29). Finally, apoD has an N-terminal glutamine residue (15), and we cannot rule out the possibility, that a covalent linkage to this residue exists and that the proteins are first cleaved by an endopeptidase thus releasing the terminal acid-Gln conjugate.…”

Section: Discussionmentioning

confidence: 87%

A Specific Bacterial Aminoacylase Cleaves Odorant Precursors Secreted in the Human Axilla

Natsch

Gfeller

Gygax

et al. 2003

Journal of Biological Chemistry

151

206

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Human axillary odor is known to be formed upon the action of Corynebacteria sp. on odorless axilla secretions. The known axilla odor determinant 3-methyl-2-hexenoic acid was identified in hydrolyzed axilla secretions along with a chemically related compound, 3-hydroxy-3-methylhexanoic acid. The natural precursors of both these acids were purified from non-hydrolyzed axilla secretions. From liquid chromatography/ mass spectrometry analysis, it appeared that the acids are covalently linked to a glutamine residue in fresh axilla secretions, and the corresponding conjugates were synthesized for confirmation. Bacterial isolates obtained from the human axilla and belonging to the Corynebacteria were found to release the acids from these odorless precursors in vitro. A Zn 2؉ -dependent aminoacylase mediating this cleavage was purified from Corynebacterium striatum Ax20, and the corresponding gene agaA was cloned and heterologously expressed in Escherichia coli. The enzyme is highly specific for the glutamine residue but has a low specificity for the acyl part of the substrate. agaA is closely related to many genes coding for enzymes involved in the cleavage of N-terminal acyl and aryl substituents from amino acids. This is the first report of the structure elucidation of precursors for human body odorants and the isolation of the bacterial enzyme involved in their cleavage.The axilla region of humans contains a dense arrangement of apocrine, eccrine, and sebaceous glands, and it is an everyday experience, that volatile substances emanating from these areas make a key contribution to human body odor. Although this odor is perceived by today's society as mainly unpleasant, several studies indicate that it may contain chemical signals that affect the menstrual cycle (1) or that may be involved in a major histocompatibility complex allele-dependent mate selection (2). These studies point to an important role of body odors in the evolutionary history of man.Sweat as it is secreted by axillary glands is odorless. Since the pioneering work of Shelley et al. (3), it is known that (a) the typical strong axilla odor can only be released from apocrine secretions, and (b) that the action of skin bacteria is needed to generate the odoriferous compounds from non-smelling molecules present in these secretions. Indeed, the axilla is a skin region supporting a dense bacterial population, which is dominated by the two genera Staphylococcus and Corynebacteria (4, 5). Most individuals carry a flora that is dominated by either one of these two genera, and a strong correlation was found between a high population of Corynebacteria and a strong axillary odor formation (4, 6). As a practical consequence of these findings, halogenated antibacterials and aluminum preparations for reducing the bacterial population have become the main active ingredient of commercial deodorants for the last 40 years. The scientific conclusion from this early work was that axilla secretions contain non-odoriferous precursors that must be transformed by bacterial en...

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“…However, the general view is that these volatile ligands need to be transported into the VNO by transport proteins belonging to the lipocalin family; in vivo, such molecules gain access to the VNO after direct contact with the stimulus source. Among these proteins that act as carrier for volatile ligands, some, such as major urinary proteins (MUPs) [15,40] or aphrodisin [17] have been now clearly characterized. It should be noticed that these carrier proteins might also serve by themselves as cues for individual recognition, especially because they exhibit a high degree of polymorphism [40] and are able to stimulate egr-1 expression in specific regions of the accessory olfactory bulb [16].…”

Section: Functional Roles Of Both the Main And The Accessory Olfactormentioning

confidence: 99%