Binding of [<sup>3</sup>H]-2-isobutyl-3-methoxypyrazine to cow olfactory mucosa

Pelosi, Paolo; Pisanelli, Anna Maria; Baldaccini, Natale Emilio; Gagliardo, Anna

doi:10.1093/chemse/6.2.77

Cited by 52 publications

(30 citation statements)

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“…Preparation of 3H-labelled 2-isobutyl-3-methoxypyrazine (1.65 Ci/mmol) was previously described [7]. All chemicals used were of reagent grade.…”

Section: Methodsmentioning

confidence: 99%

“…This latter compound has interesting properties for being a good probe for olfactory binding proteins since it has a very low olfactory threshold in humans ( 2 parts/1012) [6] and specific anosmia can be demonstrated indicating the presence of binding proteins of high affinity to 'green' odorants. The binding affinity (Kd 1.2 pM) and ligand specificity of this proteins in cows are now well characterised [5,7,IS].…”

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confidence: 99%

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Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Bignetti

Cavaggioni²,

Pelosi

et al. 1985

European Journal of Biochemistry

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158

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Cow nasal tissue contains a protein which shows specific binding activity for ‘green’ smelling compounds such as 2‐isobutyl‐3‐methoxypyrazine. This protein has now been purified using anion‐exchange fast protein liquid chromatography. The protein has a relative molecular mass of 400000–44000, s= 3.1 ± 0.3 S, pI = 4.7 ± 0.1 with an absorbance maximum at 278 nm, and consists of two subunits with an identical relative molecular mass of 19000. It is localised in the soluble fraction of cells from the olfactory mucosa and respiratory mucosa from the middle part of the maxillary and nasal turbinates, and is absent from all other tissues tested.

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“…Preparation of 3H-labelled 2-isobutyl-3-methoxypyrazine (1.65 Ci/mmol) was previously described [7]. All chemicals used were of reagent grade.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Bignetti

Cavaggioni²,

Pelosi

et al. 1985

European Journal of Biochemistry

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158

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“…The same uncertainty holds for the molecular characteristics of the complement to the odor stimulus, i.e., the odor receptor or, better stated, the reception processes. Biochemical studies have revealed the existence of an olfactory marker protein (OMP) (39,40), an odorant-binding protein (OBP) (6,48,49), and an olfactory specific G protein (G olf ) (35). The role of these proteins as well as that of adenylate cyclase as a possible second messenger in the olfactory transduction processes (46) is still the subject of study.…”

Section: Introductionmentioning

confidence: 99%

Thresholds for odor and nasal pungency

Cometto‐Muñiz

Cain

1990

Physiology & Behavior

174

137

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Detection thresholds were measured repeatedly for 11 chemicals in normosmic and anosmic subjects. The stimuli comprised the first eight members of the series of naliphatic alcohols, phenyl ethyl alcohol, pyridine, and menthol. Results showed that anosmics could detect, via pungency, all but phenyl ethyl alcohol reliably. In the aliphatic series, both odor and pungency thresholds declined with chain length in a way that implied dependence of both in part on phase distribution in the mucosa. Odor thresholds, however, declined more rapidly than pungency thresholds: the ratio of anosmics threshold/normosmics threshold increased from 23 for methanol to 10,000 for 1-octanol. The outcome of a scaling experiment employing normosmic subjects indicated that, with the exception of methanol and ethanol, pungency arose when perceived intensity reached a narrowly tuned criterion level. When thresholds were expressed as percentages of saturated vapor -an index of thermodynamic activity -thereby accounting for differences in solubility and in phase distribution in the mucosa among the various stimuli, both odor and pungency thresholds depicted a striking constancy across stimuli.

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“…In some of these instances, the findings have not been reproducible (10); in other cases, the odorant specificity and tissue distribution of the binding sites are insufficient to judge whether a specific olfactory receptor has been labeled. Pelosi and co-workers (11,12) Preparation of Bovine Tissue. Cow nasal epithelia were obtained from a local slaughterhouse within 2 hr of killing.…”

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confidence: 99%

Isolation and characterization of an olfactory receptor protein for odorant pyrazines.

Pevsner¹,

Trifiletti²,

Strittmatter³

et al. 1985

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

158

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The highly potent bell pepper odorant 2-isobutyl-3-[3H]methoxypyrazine ([3HJIBMP) binds specifically and saturably to bovine and rat nasal epithelium. Specific binding is not detected in 11 other tissues assayed, and in the rat binding is 9 times higher in olfactory than in respiratory epithelium. We have purified to apparent homogeneity a soluble pyrazine odorant binding protein that constitutes ;1% of the total soluble protein in bovine nasal epithelium. Preparation of Bovine Tissue. Cow nasal epithelia were obtained from a local slaughterhouse within 2 hr of killing. Nasal tissue was dissected from the cribriform plate to a point three-fourths of the distance to the external nares. This dissection included primary olfactory sensory neurons and respiratory tissues. Typically, dissection of a single cow nose yielded 50 g of tissue (wet weight, excluding cartilage and bone) containing 1.2 g of protein.The tissue was suspended in 2 vol of ice-cold 50 mM Tris buffer (pH 7.7 at 250C), minced finely with scissors, homogenized in a Waring blender for 30 sec, and homogenized in a Brinkmann Polytron for 30 sec. For studies on crude preparations, the suspension was centrifuged at 40,000 x g for 20 min at 4TC, and the supernatant was assayed directly. For purification studies, the suspension was centrifuged at 20,000 x g for 10 min at 4TC. The pellet was resuspended in Tris buffer, recentrifuged, and the supernatant fractions were pooled.Binding 3050The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Binding of [³H]-2-isobutyl-3-methoxypyrazine to cow olfactory mucosa

Cited by 52 publications

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Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Thresholds for odor and nasal pungency

Isolation and characterization of an olfactory receptor protein for odorant pyrazines.

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Binding of [3H]-2-isobutyl-3-methoxypyrazine to cow olfactory mucosa

Cited by 52 publications

References 0 publications

Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Purification and characterisation of an odorant‐binding protein from cow nasal tissue

Thresholds for odor and nasal pungency

Isolation and characterization of an olfactory receptor protein for odorant pyrazines.

Contact Info

Product

Resources

About

Binding of [³H]-2-isobutyl-3-methoxypyrazine to cow olfactory mucosa