Solubilization,characterization and photoaffinity labeling of the mitochondrial dihydropyridine receptor from bovine adrenal medulla |
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Institution: | 1. Qatar University, Doha 2713, Qatar;2. Pontifical Catholic University of Argentina;3. CONICET, Av. Alicia Moreau de Justo 1300, C1107AAZ, Buenos Aires, Argentina;4. International School of Management, 17, boulevard Raspail 75007 Paris France |
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Abstract: | - 1.1. The mitochondrial dihydropyridine receptor was solubilized with Chaps at a detergent/ protein ratio of 2.5, during 45 min at 4°C.
- 2.2. From the rate constants of association (8.10 ± 0.25 × 104 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.
- 3.4. The solubilized receptors are heat-resistant, sensitive to the trypsin and to the reduction of disulfide bonds.
- 4.5. In native membranes, a polypeptide of 50 kDa was specifically photolabelled with 3H]Azidopine.
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