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-rENaC
1 Departments of Anesthesiology, 2 Physiology and Biophysics, and 3 Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama 35233
We examined the effect of peroxynitrite
(ONOO
) on the cloned rat
epithelial Na+ channel
(
-rENaC) expressed in Xenopus
oocytes. 3-Morpholinosydnonimine (SIN-1) was used to concurrently
generate nitric oxide (· NO) and superoxide
(O2 ·), which react to
form ONOO, a species known
to promote protein nitration and oxidation. Under control conditions,
oocytes displayed an amiloride-sensitive whole cell conductance of 7.4 ± 2.8 (SE) µS. When incubated at 18°C with SIN-1 (1 mM) for 2 h (final ONOO concentration = 10 µM), the amiloride-sensitive conductance was reduced to
0.8 ± 0.5 µS. To evaluate whether the observed inhibition was due to ONOO, as opposed
to · NO, we also exposed oocytes to SIN-1 in the presence of
urate (500 µM), a scavenger of
ONOO and superoxide
dismutase, which scavenges
O2 ·, converting SIN-1
from an ONOO to an
· NO donor. Under these conditions, conductance values remained at control levels following SIN-1 treatment.
Tetranitromethane, an agent that oxidizes sulfhydryl groups at pH
6, also inhibited the amiloride-sensitive conductance. These data
suggest that oxidation of critical sulfhydryl groups within rENaC by
ONOO directly inhibits
channel activity.
nitric oxide; reactive species; sodium conductance; tetranitromethane; 3-morpholinosydnonimine; oxidation; nitration
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