Klotho up-regulates renal calcium channel transient receptor potential vanilloid 5 (TRPV5) by intra- And extracellular N-glycosylation-dependent mechanisms

Matthias T F Wolf, Sung Wan An, Mingzhu Nie, Manjot S. Bal, Chou Long Huang

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

The anti-aging protein Klotho is a type 1 membrane protein produced redominantly in the distal convoluted tubule. The ectodomain of Klotho is cleaved and secreted into the urine to regulate several ion channels and transporters. Secreted Klotho (sKL) up-regulates the TRPV5 calcium channel from the cell exterior by removing sialic acids from N-glycan of the channel and inhibiting its endocytosis. BecauseTRPV5and Klotho coexpress in the distal convoluted tubule, we investigated whether Klotho regulates TRPV5 action from inside the cell. Whole-cell TRPV5-mediated channel activity was recorded in HEK cells coexpressing TRPV5 and sKL or membranous Klotho (mKL). Transfection of sKL, but not mKL, produced detectable Klotho protein in cell culture media. As for sKL, mKL increased TRPV5 current density. The role of sialidase activity of mKL acting inside is supported by findings that mutations of putative sialidase activity sites in sKL and mKL abrogated the regulation of TRPV5 but that the extracellular application of a sialidase inhibitor prevented the regulation of TRPV5 by sKL only. Mechanistically, coexpression with a dominant-negative dynamin II prevented the regulation of TRPV5 by sKL but not by mKL. In contrast, blocking forward trafficking by brefeldin A prevented the effect withmKLbut not with sKL. Therefore, Klotho up-regulates TRPV5 from both the inside and outside of cells. The intracellular action of Klotho is likely due to enhanced forward trafficking of channel proteins, whereas the extracellular action is due to inhibition of endocytosis. Both effects involve putative Klotho sialidase activity. These effects of Klotho may play important roles regarding calcium reabsorption in the kidney.

Original languageEnglish (US)
Pages (from-to)35849-35857
Number of pages9
JournalJournal of Biological Chemistry
Volume289
Issue number52
DOIs
StatePublished - Dec 26 2014

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Glycosylation
TRPV Cation Channels
Calcium Channels
Up-Regulation
Kidney
Neuraminidase
Endocytosis
Dynamin II
Sialic Acids
Brefeldin A
Protein Transport
Ion Channels
Cell culture
Transfection
Polysaccharides
Culture Media
Membrane Proteins
Current density
Cell Culture Techniques
Aging of materials

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Klotho up-regulates renal calcium channel transient receptor potential vanilloid 5 (TRPV5) by intra- And extracellular N-glycosylation-dependent mechanisms. / Wolf, Matthias T F; An, Sung Wan; Nie, Mingzhu; Bal, Manjot S.; Huang, Chou Long.

In: Journal of Biological Chemistry, Vol. 289, No. 52, 26.12.2014, p. 35849-35857.

Research output: Contribution to journalArticle

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