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Am J Physiol Renal Physiol (July 30, 2002). doi:10.1152/ajprenal.00039.2002
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Articles in PresS, published online ahead of print July 30, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00039.2002
Submitted on January 29, 2002
Accepted on July 26, 2002

Activation of D2-like Receptors Causes Recruitment of Tyrosine Phosphorylated Na+, K+, ATPase {alpha}1-subunits In Kidney

Vihang A. Narkar1, Tahir Hussain1, and Mustafa F. Lokhandwala1*

1 Institute for Cardiovascular Studies College of Pharmacy, University of Houston, Houston, TX, USA

* To whom correspondence should be addressed. E-mail: MLokhandwala{at}uh.edu.

The present study investigates the cellular mechanisms responsible for dopamine D2-like receptor-mediated stimulation of Na+, K+, ATPase in the proximal tubules of the kidney. Previously, we showed that D2-like receptor-mediated increase in Na+, K+, ATPase involves an increase in the maximum rate of Na+, K+, ATPase activity (Vmax). Therefore, we tested the hypothesis that D2-like receptor-mediated stimulation of Na+, K+, ATPase requires phosphorylation and recruitment of {alpha}1-subunits of the enzyme from cytosol to the membrane. This hypothesis was tested by western blotting for Na+, K+, ATPase {alpha}1-subunits in proximal tubular membrane. Treatment of the proximal tubules with bromocriptine (D2-like receptor agonist) caused an increase in Na+, K+, ATPase {alpha}1-subunit abundance in the membrane preparations. This effect was blocked by genistein (tyrosine kinase inhibitor), suggesting a role for tyrosine phosphorylation. Moreover, bromocriptine caused an increase in tyrosine phosphorylation of membrane bound Na+, K+, ATPase {alpha}1-subunits. This effect was blocked by bafilomycin A1 (vesicular transport inhibitor), which suggested that this increase was due to the recruitment of tyrosine phosphorylated Na+, K+, ATPase {alpha}1-subunits. In conclusion, we have demonstrated that activation D2-like receptors increases Na+, K+, ATPase activity by recruitment of the tyrosine phosphorylated [[alpha1]]-subunits in the proximal tubules of the kidney.




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