title: Proton inhibition of [3H]resiniferatoxin binding to vanilloid (capsaicin) receptors in rat spinal cord.
abstract: Protons and capsaicin activate overlapping subsets of sensory nerves by opening ion conductances of similar properties. We have used the [3H]resiniferatoxin binding assay utilizing rat spinal cord membranes to elucidate the possible interaction of protons at the vanilloid (capsaicin) receptor. Using low pH (pH 6.0 and pH 5.0) buffers, a time-dependent gradual decrease was observed in specific resiniferatoxin binding. Protons inhibited resiniferatoxin binding with an IC50 of pH 5.3 +/- 0.1. In experiments in which the concentration of [3H]resiniferatoxin was varied, protons reduced the Bmax value by approximately 40% with a corresponding 2-fold decrease in affinity. No change however, was observed in binding cooperativity (the Hill coefficients were 1.7 +/- 0.1 and 1.6 +/- 0.2 in the presence of pH 7.4 and pH 5.0 buffers, respectively). These changes in binding parameters are consistent with a non-competitive or, alternatively, mixed inhibitory mechanism. The remaining resiniferatoxin binding sites bound capsaicin with an affinity (Ki = 5.0 +/- 1.0 microM) very similar to that determined in the presence of a pH 7.4 buffer (Ki = 3.0 +/- 1.5 microM). A cyclooxygenase inhibitor, indomethacin (up to 10 microM), did not prevent the action of protons on resiniferatoxin binding; neither was it mimicked by prostanoids (prostaglandin I2 and E1, both at 100 microM). We conclude that protons interact at vanilloid receptors in the rat spinal cord; this interaction is either non-competitive or mixed in nature, and probably is not related to prostanoid generation. Protons and/or putative proton-generated mediators might represent endogenous modulators of the vanilloid receptor.
authors.affiliation: Department of Physiology and Pharmacology, Karolinska Institute, Stockholm, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Szallasi
authors.forename: P M
authors.initials: PM
authors.lastname: Blumberg
authors.forename: J M
authors.initials: JM
authors.lastname: Lundberg
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002211:Capsaicin / Q000494:pharmacology*; D004224:Diterpenes / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006863:Hydrogen-Ion Concentration; D011522:Protons; D051381:Rats; D017207:Rats, Sprague-Dawley; D011955:Receptors, Drug / Q000187:drug effects* / Q000378:metabolism; D013116:Spinal Cord / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors
chemical_list: D004224:Diterpenes; D011522:Protons; D011955:Receptors, Drug; C024353:resiniferatoxin; D002211:Capsaicin