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Biophysical Journal 69: 106-119 (1995)
© 1995 the Biophysical Society
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill 27599, USA.
ABSTRACT
The calmodulin-binding properties of the rabbit skeletal muscle Ca2+ release channel (ryanodine receptor) and the channel's regulation by calmodulin were determined at < or = 0.1 microM and micromolar to millimolar Ca2+ concentrations. [125I]Calmodulin and [3H]ryanodine binding to sarcoplasmic reticulum (SR) vesicles and purified Ca2+ release channel preparations indicated that the large (2200 kDa) Ca2+ release channel complex binds with high affinity (KD = 5-25 nM) 16 calmodulins at < or = 0.1 microM Ca2+ and 4 calmodulins at 100 microM Ca2+. Calmodulin-binding affinity to the channel showed a broad maximum at pH 6.8 and was highest at 0.15 M KCl at both < or = 0.1 MicroM and 100 microM Ca2+. Under condition closely related to those during muscle contraction and relaxation, the half-times of calmodulin dissociation and binding were 50 +/- 20 s and 30 +/- 10 min, respectively. SR vesicle-45Ca2+ flux, single-channel, and [3H]ryanodine bind measurements showed that, at < or = 0.2 microM Ca2+, calmodulin activated the Ca2+ release channel severalfold. Ar micromolar to millimolar Ca2+ concentrations, calmodulin inhibited the Ca(2+)-activated channel severalfold. Hill coefficients of approximately 1.3 suggested no or only weak cooperative activation and inhibition of Ca2+ release channel activity by calmodulin. These results suggest a role for calmodulin in modulating SR Ca2+ release in skeletal muscle at both resting and elevated Ca2+ concentrations.
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S. Sencer, R. V. L. Papineni, D. B. Halling, P. Pate, J. Krol, J.-Z. Zhang, and S. L. Hamilton Coupling of RYR1 and L-type Calcium Channels via Calmodulin Binding Domains J. Biol. Chem., October 5, 2001; 276(41): 38237 - 38241. [Abstract] [Full Text] [PDF]
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G. G. Rodney, C. P. Moore, B. Y. Williams, J.-Z. Zhang, J. Krol, S. E. Pedersen, and S. L. Hamilton Calcium Binding to Calmodulin Leads to an N-terminal Shift in Its Binding Site on the Ryanodine Receptor J. Biol. Chem., January 12, 2001; 276(3): 2069 - 2074. [Abstract] [Full Text] [PDF]
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D. M. Balshaw, L. Xu, N. Yamaguchi, D. A. Pasek, and G. Meissner Calmodulin Binding and Inhibition of Cardiac Muscle Calcium Release Channel (Ryanodine Receptor) J. Biol. Chem., June 1, 2001; 276(23): 20144 - 20153. [Abstract] [Full Text] [PDF]
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