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-rENaC alter sensitivity to amiloride and reactive species
1 Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, AL, USA
2 Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
3 Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
* To whom correspondence should be addressed. E-mail: sadis{at}uab.edu.
We studied the effects of two mutations of the extracellular loop of 
rENaC on amiloride-sensitive current in Xenopus oocytes and the inhibition of this current by SIN- 1. Injection of oocytes with wild type (wt) ,
,
-rENaC cRNA (8.3 ng per subunit)
resulted 48-72 h later in inward Na+ currents (-5.5 ± 0.8 µA; mean ± SEM at -100 mV;
n=21) which were completely inhibited by amiloride. Oocytes injected with either Y279A
or Y283A and ,-rENaC cRNAs had significantly lower Na+ currents. Furthermore Y279A ,-rENaC injected oocytes had a higher Ki for amiloride (0.54±0.97 vs. 0.10±0.04 µM; p<0.01). Exposure of oocytes to SIN-1 (1 mM) for five minutes
decreased both total Na+ and amiloride-sensitive currents across wt- and Y279A but not
Y283A ,-rENaC. Furthermore, exposure to SIN-1 increased the Ki for amiloride across wt but not Y279A ,-rENaC injected oocytes. These data indicate that both tyrosines are important for proper ENaC function and their oxidative modifications contribute to
altered ENaC function.
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