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1 Department of Physiology, Nagoya City University Medical School, Nagoya 467-8601; and 2 Department of Pharmacology, Juntendo University School of Medicine, Tokyo 113-8421, Japan
The type 1 ryanodine
receptor (RyR1) from rabbit skeletal muscle displayed two distinct
degrees of response to cytoplasmic Ca2+ [high- and
low-open probability (Po) channels]. Here, we
examined the effects of adenine nucleotides and caffeine on these
channels and their modulations by sulfhydryl reagents.
High-Po channels showed biphasic
Ca2+ dependence and were activated by adenine nucleotides
and caffeine. Unexpectedly, low-Po channels did
not respond to either modulator. The addition of a reducing reagent,
dithiothreitol, to the cis side converted the
high-Po channel to a state similar to that of
the low-Po channel. Treatment with
p-chloromercuriphenylsulfonic acid (pCMPS) transformed
low-Po channels to a
high-Po channel-like state with stimulation by
,
-methylene-ATP and caffeine. In experiments under redox control
using glutathione buffers, shift of the cis potential toward
the oxidative state activated the low-Po
channel, similar to that of the high-Po or the
pCMPS-treated channel, whereas reductive changes inactivated the
high-Po channel. Changes in trans
redox potential, in contrast, did not affect channel activity of
either channel. In all experiments, channels with higher
Po were stimulated to a great extent by
modulators, but ones with lower Po were
unresponsive. These results suggest that redox states of critical
sulfhydryls located on the cytoplasmic side of the RyR1 may alter both
gating properties of the channel and responsiveness to channel modulators.
calcium-induced calcium release; adenine nucleotide; caffeine; sulfhydryl reagents; redox potential
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