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Angiotensin‐II induced protein kinase D activation via src kinase and protein kinase C mediates aldosterone secretion in bovine adrenal glomerulosa cells
Author(s) -
Olala Lawrence Ogweno,
Bollag Wendy B.
Publication year - 2010
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.24.1_supplement.lb599
Subject(s) - aldosterone , protein kinase c , angiotensin ii , endocrinology , medicine , tyrosine kinase , zona glomerulosa , proto oncogene tyrosine protein kinase src , protein kinase a , kinase , phosphorylation , secretion , biology , chemistry , microbiology and biotechnology , receptor , blood pressure
Objective To study the mechanism of angiotensin II (AngII) induced protein kinase D (PKD) activation in modulating aldosterone secretion in bovine adrenal glomerulosa (BAG) cells. Methods Adenoviral vectors were used to overexpress S738/742A PKD mutant which is not phosphorylatable by protein kinase C (PKC), Y463F PKD mutant, and Src wildtype as well as a dominant negative K296R/Y528F Src mutant. Supernatants were stored for aldosterone assay, and the cells processed for immunoblot analysis. Samples were immunoprecipitated with either PKD, or anti‐phosphotyrosine antibodies and reciprocally blotted. Blots were quantified and statistically significant differences were determined when p<0.05. Results Overexpression of non‐phosphorylatable S738/742A and the Y463F PKD mutants downregulated PKD activation and decreased acute (30‐minute) aldosterone secretion. The dominant negative K296R/Y528F Src mutant, which blocks Src activation, attenuated PKD phosphorylation and aldosterone secretion. AngII induced an increase in tyrosine phosphorylation of PKD presumably at the tyrosine 463 residue. Conclusion Angiotensin II (AngII) activates protein kinase D in BAG cells via a mechanism involving Src kinase and PKC, to underlie increased aldosterone production. Funding Support National Institutes of Health/National Heart, Lung and Blood Institute American Heart Association