{"title":"[3H]鱼毒素A-20-α-苯甲酸酯和[3H]蛤毒素与鳟鱼脑突触体钠通道相关受体位点的结合","authors":"Jared G. Rubin, David M. Soderlund","doi":"10.1016/0742-8413(93)90200-5","DOIUrl":null,"url":null,"abstract":"<div><p>1. [<sup>3</sup>H]Batrachotoxinin A-20-α-benzoate ([<sup>3</sup>H]BTX-b) and [<sup>3</sup>H]saxitoxin ([<sup>3</sup>H]STX), radioligands that bind to distinct sites on the voltage-sensitive sodium channel, were bound specifically to saturable sites in rainbow trout (<em>Oncorhynchus mykiss</em>) brain synaptoneurosomes.</p><p>2. Specific [<sup>3</sup>H]BTX-B binding was temperature dependent with highest levels of specific [<sup>3</sup>H]BTX-B binding observed at 7°C. Specific binding was inversely correlated with assay temperature at temperatures above 7°C.</p><p>3. Saturating concentrations of scorpion (<em>Leiurus quinquestriatus</em>) venom (ScV) stimulated specific [<sup>3</sup>H]BTX-B binding at 27°C, but not at 7°C. The dihydropyrazole insecticide RH 3421 inhibited specific [<sup>3</sup>H]BTX-B binding at 7°C but had no effect on specific binding at 27°C. The sodium channel activators veratridine and aconitine and the local anesthetic dibucaine inhibited specific [<sup>3</sup>H]BTX-B binding at both 7°C and 27°C.</p><p>4. Displacement experiments in the presence of ScV at 27°C gave an equilibrium dissociation constant (<em>K</em><sub><span>d</span></sub>) for [<sup>3</sup>H]BTX-B of 710 nM and a maximal binding capacity (<em>B</em><sub>max</sub>) of 11.3 pmol/mg protein. Kinetic experiments established the rates of association (1.17 × 10<sup>5</sup>min<sup>−1</sup> nM<sup>−1</sup>) and dissociation (0.0514min<sup>−1</sup>) of the ligand-receptor complex.</p><p>5. The binding of [<sup>3</sup>H]STX reached apparent saturation at 7.5 nM. Scatchard analysis of the saturation data indicated a <em>K</em><sub><span>d</span></sub> of 3.8nM and a <em>B</em><sub>max</sub> of 1.9 pmol/mg protein.</p><p>6. These studies provide evidence for high affinity, saturable binding sites for [<sup>3</sup>H]BTX-B and [<sup>3</sup>H]STX in trout brain preparations. Whereas certain neurotoxins modified the specific binding of [<sup>3</sup>H]BTX-B in trout brain synaptoneurosomes in a predictable fashion, other compounds known to affect specific [<sup>3</sup>H]BTX-B binding in mammalian brain preparations had no effect on specific [<sup>3</sup>H]BTX-B binding in the trout.</p></div>","PeriodicalId":72650,"journal":{"name":"Comparative biochemistry and physiology. C: Comparative pharmacology","volume":"105 2","pages":"Pages 231-238"},"PeriodicalIF":0.0000,"publicationDate":"1993-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0742-8413(93)90200-5","citationCount":"5","resultStr":"{\"title\":\"Binding of [3H]batrachotoxinin A-20-α-benzoate and [3h]saxitoxin to receptor sites associated with sodium channels in trout brain synaptoneurosomes\",\"authors\":\"Jared G. Rubin, David M. Soderlund\",\"doi\":\"10.1016/0742-8413(93)90200-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>1. [<sup>3</sup>H]Batrachotoxinin A-20-α-benzoate ([<sup>3</sup>H]BTX-b) and [<sup>3</sup>H]saxitoxin ([<sup>3</sup>H]STX), radioligands that bind to distinct sites on the voltage-sensitive sodium channel, were bound specifically to saturable sites in rainbow trout (<em>Oncorhynchus mykiss</em>) brain synaptoneurosomes.</p><p>2. Specific [<sup>3</sup>H]BTX-B binding was temperature dependent with highest levels of specific [<sup>3</sup>H]BTX-B binding observed at 7°C. Specific binding was inversely correlated with assay temperature at temperatures above 7°C.</p><p>3. Saturating concentrations of scorpion (<em>Leiurus quinquestriatus</em>) venom (ScV) stimulated specific [<sup>3</sup>H]BTX-B binding at 27°C, but not at 7°C. The dihydropyrazole insecticide RH 3421 inhibited specific [<sup>3</sup>H]BTX-B binding at 7°C but had no effect on specific binding at 27°C. The sodium channel activators veratridine and aconitine and the local anesthetic dibucaine inhibited specific [<sup>3</sup>H]BTX-B binding at both 7°C and 27°C.</p><p>4. Displacement experiments in the presence of ScV at 27°C gave an equilibrium dissociation constant (<em>K</em><sub><span>d</span></sub>) for [<sup>3</sup>H]BTX-B of 710 nM and a maximal binding capacity (<em>B</em><sub>max</sub>) of 11.3 pmol/mg protein. Kinetic experiments established the rates of association (1.17 × 10<sup>5</sup>min<sup>−1</sup> nM<sup>−1</sup>) and dissociation (0.0514min<sup>−1</sup>) of the ligand-receptor complex.</p><p>5. The binding of [<sup>3</sup>H]STX reached apparent saturation at 7.5 nM. Scatchard analysis of the saturation data indicated a <em>K</em><sub><span>d</span></sub> of 3.8nM and a <em>B</em><sub>max</sub> of 1.9 pmol/mg protein.</p><p>6. These studies provide evidence for high affinity, saturable binding sites for [<sup>3</sup>H]BTX-B and [<sup>3</sup>H]STX in trout brain preparations. Whereas certain neurotoxins modified the specific binding of [<sup>3</sup>H]BTX-B in trout brain synaptoneurosomes in a predictable fashion, other compounds known to affect specific [<sup>3</sup>H]BTX-B binding in mammalian brain preparations had no effect on specific [<sup>3</sup>H]BTX-B binding in the trout.</p></div>\",\"PeriodicalId\":72650,\"journal\":{\"name\":\"Comparative biochemistry and physiology. C: Comparative pharmacology\",\"volume\":\"105 2\",\"pages\":\"Pages 231-238\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1993-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/0742-8413(93)90200-5\",\"citationCount\":\"5\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Comparative biochemistry and physiology. C: Comparative pharmacology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/0742841393902005\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Comparative biochemistry and physiology. C: Comparative pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/0742841393902005","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Binding of [3H]batrachotoxinin A-20-α-benzoate and [3h]saxitoxin to receptor sites associated with sodium channels in trout brain synaptoneurosomes
1. [3H]Batrachotoxinin A-20-α-benzoate ([3H]BTX-b) and [3H]saxitoxin ([3H]STX), radioligands that bind to distinct sites on the voltage-sensitive sodium channel, were bound specifically to saturable sites in rainbow trout (Oncorhynchus mykiss) brain synaptoneurosomes.
2. Specific [3H]BTX-B binding was temperature dependent with highest levels of specific [3H]BTX-B binding observed at 7°C. Specific binding was inversely correlated with assay temperature at temperatures above 7°C.
3. Saturating concentrations of scorpion (Leiurus quinquestriatus) venom (ScV) stimulated specific [3H]BTX-B binding at 27°C, but not at 7°C. The dihydropyrazole insecticide RH 3421 inhibited specific [3H]BTX-B binding at 7°C but had no effect on specific binding at 27°C. The sodium channel activators veratridine and aconitine and the local anesthetic dibucaine inhibited specific [3H]BTX-B binding at both 7°C and 27°C.
4. Displacement experiments in the presence of ScV at 27°C gave an equilibrium dissociation constant (Kd) for [3H]BTX-B of 710 nM and a maximal binding capacity (Bmax) of 11.3 pmol/mg protein. Kinetic experiments established the rates of association (1.17 × 105min−1 nM−1) and dissociation (0.0514min−1) of the ligand-receptor complex.
5. The binding of [3H]STX reached apparent saturation at 7.5 nM. Scatchard analysis of the saturation data indicated a Kd of 3.8nM and a Bmax of 1.9 pmol/mg protein.
6. These studies provide evidence for high affinity, saturable binding sites for [3H]BTX-B and [3H]STX in trout brain preparations. Whereas certain neurotoxins modified the specific binding of [3H]BTX-B in trout brain synaptoneurosomes in a predictable fashion, other compounds known to affect specific [3H]BTX-B binding in mammalian brain preparations had no effect on specific [3H]BTX-B binding in the trout.
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