Parts of this study were presented at the 66th and 67th Regional Meetings (Kinki Area) of the Japanese Pharmacological Society (Fukui et al., 1985; Takemura et al., 1985).
Solubilization and Characterization of the Nitrendipine Receptor in Rat Brain
Version of Record online: 5 OCT 2006
Journal of Neurochemistry
Volume 48, Issue 4, pages 1185–1190, April 1987
How to Cite
Takemura, M., Fukui, H., Yamamoto, Y., Hosomi, N. and Wada, H. (1987), Solubilization and Characterization of the Nitrendipine Receptor in Rat Brain. Journal of Neurochemistry, 48: 1185–1190. doi: 10.1111/j.1471-4159.1987.tb05645.x
- Issue online: 5 OCT 2006
- Version of Record online: 5 OCT 2006
- Received November 27, 1985; accepted October 15, 1986
- Calcium channel;
- Gel filtration;
- Sucrose density gradient sedimentation;
- Molecular weight)
Abstract: The nitrendipine receptor associated with the voltage-dependent calcium channel in rat brain was solubilized by detergent extraction and sonication. The detergent solution used for extraction consisted of 10 mM 3-[(3-cholamidopropyl)dimethylammonio] - 1 - propanesulfonate (CHAPS), 0.25% (wt/vol) polyoxyethylene 20 cetyl ether (Brij 58), and 0.025% (wt/vol) polyoxyethylene 17 cetyl stearyl ether (Lubrol WX) in the presence of 30% (wt/vol) glycerol as a stabilizer. The molecular weight of the receptor was estimated to be 1,800K by Sephacryl S-500 gel filtration and 800K by sucrose density gradient sedimentation. The equilibrium dissociation constant of [3H]nitrendipine to the solubilized receptors was 5.6 nM, which is ∼10 times that of the membrane-bound receptor. The binding of nitrendipine to the receptor was inhibited noncompetitively by the structurally unrelated calcium channel inhibitors verapamil and prenylamine; their concentrations for 50% inhibition were both 1.0 ∼ 10−7M, and they caused maximal inhibitions of 70 and 100%, respectively.