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Role of the putative PKC phosphorylation sites of the type IIc sodium-dependent phosphate transporter in parathyroid hormone regulation

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Abstract

Background

Injection of parathyroid hormone (PTH) rapidly stimulates renal Pi excretion, in part by downregulating NaPi-IIa (Npt2a/SLC34A1) and NaPi-IIc (Npt2c/SLC34A3) transporters. The mechanisms underlying the effects of PTH on NaPi-IIc are not fully elucidated.

Methods

We analyzed the effect of PTH on inorganic phosphate (Pi) reabsorption in Npt2a−/− mice to eliminate the influence of Npt2a on renal Pi reabsorption. In opossum kidney (OK) cells and Xenopus oocytes, we investigated the effect of NaPi-IIc transporter phosphorylation. Studies of mice with mutations of NaPi-IIc protein in which serine and threonine were replaced with either alanine (A), which prevents phosphorylation, or aspartic acid (D), which mimics the charged state of phosphorylated NaPi-IIc, were also performed to evaluate the involvement of phosphorylation in the regulation of transport function.

Results

The Npt2a−/− experiments showed that PTH administration rapidly inactivated NaPi-IIc function in the apical membrane of proximal tubular cells. Analysis of mutant proteins (S71, S138, T151, S174, T583) at putative protein kinase C sites, revealed that S138 markedly suppressed the function and cellular expression of mouse NaPi-IIc in Xenopus oocytes and OK cells. In addition, 138D had a short half-life compared with wild-type protein.

Conclusions

The present study suggests that acute regulation of NaPi-IIc protein by PTH is involved in the inactivation of Na+-dependent Pi cotransporter activity and that phosphorylation of the transporter is involved in the rapid modification.

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Acknowledgements

This study was supported by Support Center for Advanced Medical Sciences, Tokushima University Graduate School of Biomedical Sciences. This study was in part supported by a grant-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (no. 23689045 to H. Segawa, no. 26293204 to K. Miyamoto), and The Salt Science Research Foundation (Japan).

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Authors and Affiliations

  1. Department of Molecular Nutrition, Institute of Biomedical Sciences, University of Tokushima Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan

    Toru Fujii, Hiroko Segawa, Ai Hanazaki, Shiori Nishiguchi, Sakura Minoshima, Akiko Ohi, Rieko Tominaga, Sumire Sasaki, Kazuya Tanifuji, Megumi Koike, Yuki Arima, Yuji Shiozaki, Ichiro Kaneko, Sawako Tatsumi & Ken-ichi Miyamoto

  2. Human Science and Environment, University of Hyogo Graduate School, Hyogo, Japan

    Mikiko Ito

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  1. Toru Fujii
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Correspondence to Hiroko Segawa or Ken-ichi Miyamoto.

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This article does not contain any studies with human performed by any of the authors. Mice were handled in accordance with the Guidelines for Animal Experimentation of Tokushima University School of Medicine (T29-3).

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Fujii, T., Segawa, H., Hanazaki, A. et al. Role of the putative PKC phosphorylation sites of the type IIc sodium-dependent phosphate transporter in parathyroid hormone regulation. Clin Exp Nephrol 23, 898–907 (2019). https://doi.org/10.1007/s10157-019-01725-6

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  • DOI: https://doi.org/10.1007/s10157-019-01725-6

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