Total [3H]Ryanodine Bound | n | |||
---|---|---|---|---|
Control | 1 mM Suramin | |||
pmol/mg HSR | ||||
None | 3.38 ± 0.35 | 7.55 ± 0.69 | 5 | |
Cold ryanodine | 100 μM | 0.051 ± 0.010 | 0.09 ± 0.02 | 5 |
Ruthenium red | 10 μM | 0.058 ± 0.011 | 0.53 ± 0.03 | 5 |
20 μM | 0.045 ± 0.004 | 0.39 ± 0.02 | 3 | |
EGTA | 5 mM | 0.046 ± 0.005 | 1.11 ± 0.05 | 3 |
10 mM | 0.044 ± 0.002 | 1.16 ± 0.01 | 2 | |
MgCl2 | 10 mM | 0.132 ± 0.025 | 5.38 ± 0.46 | 5 |
30 mM | 0.077 ± 0.013 | 4.69 ± 0.45 | 5 | |
50 mM | 0.066 ± 0.012 | 4.50 ± 0.44 | 5 | |
CaCl2 | 10 mM | 0.161 ± 0.030 | 5.64 ± 0.47 | 5 |
30 mM | 0.084 ± 0.007 | 4.93 ± 0.46 | 5 | |
50 mM | 0.061 ± 0.008 | 4.02 ± 0.41 | 5 | |
4-CMPS | 100 μM | 0.262 ± 0.062 | 0.31 ± 0.08 | 5 |
200 μM | 0.204 ± 0.042 | 0.22 ± 0.05 | 5 | |
Neomycin | 1 mM | 0.680 ± 0.012 | 7.07 ± 0.56 | 5 |
Tetracaine | 1 mM | 0.265 ± 0.046 | 5.46 ± 0.41 | 3 |
[3H]Ryanodine binding was performed in the absence or presence of 1 mM suramin with 40 mM Mops/Tris, pH 7.0, 10 nM [3H]ryanodine, 0.5 M CsCl, protease inhibitors (seeExperimental Procedures) and the indicated agents at 37°C for 90 min. Values are presented as means ± S.E.M. of total [3H]ryanodine binding from five experiments with different HSR preparations.