Identification of calcium-calmodulin multifunctional protein kinase II in rabbit kidney

Recent studies have demonstrated that calcium/calmodulin-dependent multifunctional protein kinase II (CaM-KII) inhibits the reconstituted Na(+)-H+ exchanger from the brush border membrane of proximal convoluted tubule of the rabbit kidney. The present studies were undertaken to evaluate the physiological relevance of this finding by establishing the presence of CaM-KII in rabbit kidney and proximal convoluted tubule cells by Northern RNA hybridization analysis to demonstrate the messenger RNA (mRNA) for CaM-KII and by a selective enzymatic assay of CaM-KII using a synthetic peptide substrate. A single 4.9 Kb mRNA was observed on hybridization of total RNA from rabbit kidney cortex and medulla and from an enriched suspension of rabbit kidney proximal convoluted tubules with a cDNA for rat brain CaM-KII. An enzyme assay using a synthetic peptide substrate representing the site phosphorylated by CaM-KII on glycogen synthase demonstrated calcium-calmodulin dependent protein kinase activity in both rabbit kidney cortex (specific activity of 662 +/- 127 nmol.min-1.mg protein-1) and proximal tubule cells (546 +/- 77 nmol.min-1.mg protein-1). These data establish the presence of CaM-KII in the rabbit kidney, and suggest a role for this enzyme in the control of renal electrolyte transport.