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Solubilization, characterization and photoaffinity labeling of the mitochondrial dihydropyridine receptor from bovine
M J Hidalgo1, L M Gutierrez, M Palmero
1Departamento de Neuroquímica, Universidad de Alicante, Spain.
The International Journal of Biochemistry
|December 1, 1993
Abstract:
1. The mitochondrial dihydropyridine receptor was solubilized with Chaps at a detergent/protein ratio of 2.5, during 45 min at 4 degrees C. 2. From the rate constants of association (8.10 +/- 0.25 x 10(4) M-1 min-1) and dissociation (0.022 +/- 0.001 min-1) a Kd of 275 nM was calculated, while from saturation experiments a Kd of 270 +/- 30 nM and a density of receptors of 106 +/- 9 pmol/mg protein was obtained. 4. The solubilized receptors are heat-resistant, sensitive to the trypsin and to the reduction of disulfide bonds. 5. In native membranes, a polypeptide of 50 kDa was specifically photolabelled with [3H]Azidopine.