Dynamic regulation of FGF23 by Fam20C phosphorylation, GalNAc-T3 glycosylation, and furin proteolysis

Proc Natl Acad Sci U S A. 2014 Apr 15;111(15):5520-5. doi: 10.1073/pnas.1402218111. Epub 2014 Mar 26.

Abstract

The family with sequence similarity 20, member C (Fam20C) has recently been identified as the Golgi casein kinase. Fam20C phosphorylates secreted proteins on Ser-x-Glu/pSer motifs and loss-of-function mutations in the kinase cause Raine syndrome, an often-fatal osteosclerotic bone dysplasia. Fam20C is potentially an upstream regulator of the phosphate-regulating hormone fibroblast growth factor 23 (FGF23), because humans with FAM20C mutations and Fam20C KO mice develop hypophosphatemia due to an increase in full-length, biologically active FGF23. However, the mechanism by which Fam20C regulates FGF23 is unknown. Here we show that Fam20C directly phosphorylates FGF23 on Ser(180), within the FGF23 R(176)XXR(179)/S(180)AE subtilisin-like proprotein convertase motif. This phosphorylation event inhibits O-glycosylation of FGF23 by polypeptide N-acetylgalactosaminyltransferase 3 (GalNAc-T3), and promotes FGF23 cleavage and inactivation by the subtilisin-like proprotein convertase furin. Collectively, our results provide a molecular mechanism by which FGF23 is dynamically regulated by phosphorylation, glycosylation, and proteolysis. Furthermore, our findings suggest that cross-talk between phosphorylation and O-glycosylation of proteins in the secretory pathway may be an important mechanism by which secreted proteins are regulated.

Keywords: Fam20; chronic kidney disease; familial tumoral calcinosis; phosphate homeostasis; rickets.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Casein Kinase I
  • DNA Primers / genetics
  • Extracellular Matrix Proteins / immunology
  • Extracellular Matrix Proteins / metabolism*
  • Fibroblast Growth Factor-23
  • Fibroblast Growth Factors / metabolism*
  • Furin / metabolism*
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Mass Spectrometry
  • Molecular Sequence Data
  • N-Acetylgalactosaminyltransferases / metabolism*
  • Phosphorylation
  • Polypeptide N-acetylgalactosaminyltransferase
  • Proteolysis
  • Sequence Analysis, DNA
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tandem Mass Spectrometry

Substances

  • DNA Primers
  • Extracellular Matrix Proteins
  • FGF23 protein, human
  • Fgf23 protein, mouse
  • Fibroblast Growth Factors
  • Fibroblast Growth Factor-23
  • N-Acetylgalactosaminyltransferases
  • Casein Kinase I
  • FAM20C protein, human
  • Furin