Expression of HKalpha2 protein is increased selectively in renal medulla by chronic hypokalemia.
Our laboratory has demonstrated by Northern analysis that chronic hypokalemia increases HKalpha2 (i.e., alpha-subunit of the colonic H+-K+-ATPase) mRNA abundance in the rat. To determine whether the increase in mRNA correlated with an increase in HKalpha2 protein, an antibody was raised against a synthetic peptide derived from amino acids 686-698 of the HKalpha2 sequence. The anti-HKalpha2 antibody hybridized to rat distal colon membranes which migrated at approximately 100 kDa (expected mobility of HKalpha2). HKalpha2 protein was not detected in plasma membranes from rat whole kidney or stomach (100 microg) derived from control animals. The antibody was then used to investigate changes in expression of HKalpha2 in renal cortex, renal medulla, and distal colon in two pathophysiological conditions: 1) chronic hypokalemia (LK) and 2) chronic metabolic acidosis (CMA). In LK rats there was a marked, but selective, increase in the abundance of HKalpha2 protein in membranes prepared from renal medulla. Nevertheless, a corresponding increase in HKalpha2 protein abundance was not observed in membranes prepared from the distal colon of LK rats. HKalpha2 protein abundance in CMA was indistinguishable from controls. Moreover, chronic hypokalemia had no effect on expression of alpha1-Na+-K+-ATPase or HKalpha1 in kidney or distal colon under any experimental condition. Therefore, HKalpha2 protein is tissue- and site-specifically upregulated in response to chronic hypokalemia but not by CMA. Furthermore, this regulatory response is localized to the renal medulla.
Codina
J
J
Division of Renal Diseases and Hypertension, Department of Internal Medicine, University of Texas Houston Medical School, Houston, Texas 77030, USA.
Delmas-Mata
J T
JT
DuBose
T D
TD
D000818:Animals; D000918:Antibody Specificity; D001921:Brain / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D003106:Colon / Q000201:enzymology; D015870:Gene Expression; D017506:H(+)-K(+)-Exchanging ATPase / Q000235:genetics* / Q000276:immunology; D007008:Hypokalemia / Q000201:enzymology*; D015151:Immunoblotting; D007527:Isoenzymes / Q000235:genetics* / Q000276:immunology; D007679:Kidney Medulla / Q000201:enzymology*; D008297:Male; D010446:Peptide Fragments / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D007527:Isoenzymes; D010446:Peptide Fragments; D012333:RNA, Messenger; D017506:H(+)-K(+)-Exchanging ATPase