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Protein Carbonylation Regulates Renal Proximal Tubular Na/K‐ATPase signaling and Sodium Transport
Author(s) -
YAN YANLING,
Xie Zijian,
Shapiro Joseph I.,
Liu Jiang
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1115.11
Subject(s) - chemistry , ouabain , proto oncogene tyrosine protein kinase src , renal sodium reabsorption , mg132 , atpase , reabsorption , signal transduction , microbiology and biotechnology , biochemistry , sodium , proteasome , biology , proteasome inhibitor , enzyme , organic chemistry
We have shown that cardiotonic steroids signaling through the Na/K‐ATPase regulate sodium reabsorption in renal proximal tubule (RPT). Here we report that protein carbonylation is critical in modulation of Na/K‐ATPase signaling and RPT ion transport. In RPT LLC‐PK1 cells, both ouabain (100nM) and glucose oxidase (GO, 1 and 3mU/ml) stimulated protein carbonylation of Na/K‐ATPase α1 subunit and c‐Src, activated c‐Src, stimulated redistribution of Na/K‐ATPase α1 and NHE3, and inhibited active transepithelial 22 Na + flux. Pretreatment with the antioxidant, N‐Acetyl‐L‐Cysteine (NAC), can prevent these effects in a dose‐dependent manner. Furthermore, disruption of the Na/K‐ATPase/c‐ Src signaling complex and c‐Src inhibitor PP2 abolished ouabain‐induced ROS generation and protein carbonylation. Moreover, it appears that there is a self‐regulatory mechanism to reverse carbonylation after removal of ouabain from culture medium, evaluated in the presence of protein biosynthesis inhibitor cycloheximide, proteasome inhibitor MG132, and lysosomotropic weak base agent chloroquine. In summary, our data indicated that protein carbonylation is involved in a feed‐forward mechanism in regulation of Na/K‐ATPase/ c‐Src signaling and related sodium transport. The data also suggested that the Na/K‐ATPase/c‐Src signaling might be a functional receptor of ROS. (Supported by NIH HL‐109015 to Z.X. and J.I.S.).