BACKGROUND/AIM: Parathyroid hormone (PTH)-dependent inhibition of proximal tubular P(i) reabsorption is mediated by protein kinase A and/or C and is associated with reduced border membrane expression of the type IIa Na/P(i) cotransporter. The aim of this study was to analyze phosphorylation of the type IIa cotransporter protein. METHODS: Opossum kidney cells were used as a 'proximal tubular' cell model. Protein phosphorylation was determined by immunoprecipitation of the type IIa Na/P(i) cotransporter, followed by autoradiography. The transporter protein content was evaluated by Western blotting and transport activity by tracer P(i) uptake. RESULTS: Under control conditions (no PTH) the transporter was phosphorylated; upon treatment with PTH, a decrease in phosphorylation was observed. A protein phosphatase inhibitor (okadaic acid) was unable to prevent PTH-induced Na/P(i) cotransporter inhibition but reduced transporter degradation. CONCLUSION: The type IIa Na/P(i) cotransporter is a phosphoprotein, but alterations in its phosphorylation seem not to be involved in P(i) transport inhibition.
BACKGROUND/AIM: Parathyroid hormone (PTH)-dependent inhibition of proximal tubular P(i) reabsorption is mediated by protein kinase A and/or C and is associated with reduced border membrane expression of the type IIa Na/P(i) cotransporter. The aim of this study was to analyze phosphorylation of the type IIa cotransporter protein. METHODS: Opossum kidney cells were used as a 'proximal tubular' cell model. Protein phosphorylation was determined by immunoprecipitation of the type IIa Na/P(i) cotransporter, followed by autoradiography. The transporter protein content was evaluated by Western blotting and transport activity by tracer P(i) uptake. RESULTS: Under control conditions (no PTH) the transporter was phosphorylated; upon treatment with PTH, a decrease in phosphorylation was observed. A protein phosphatase inhibitor (okadaic acid) was unable to prevent PTH-induced Na/P(i) cotransporter inhibition but reduced transporter degradation. CONCLUSION: The type IIa Na/P(i) cotransporter is a phosphoprotein, but alterations in its phosphorylation seem not to be involved in P(i) transport inhibition.
Authors: So Nagai; Makoto Okazaki; Hiroko Segawa; Clemens Bergwitz; Thomas Dean; John T Potts; Matthew J Mahon; Thomas J Gardella; Harald Jüppner Journal: J Biol Chem Date: 2010-11-03 Impact factor: 5.157