Low stability and a conserved N-glycosylation site are associated with regulation of the discoidin domain receptor family by glucose via post-translational N-glycosylation

Biosci Biotechnol Biochem. 2013;77(9):1907-16. doi: 10.1271/bbb.130351. Epub 2013 Sep 7.

Abstract

Cell-surface expression of the discoidin domain receptor (DDR) tyrosine kinase family in high molecular mass form was controlled sensitively by the glucose concentration through a post-translational N-glycosylation process. Cycloheximide time-course experiments revealed that the high-molecular-mass forms of DDR1 and DDR2 were significantly less stable than control receptor tyrosine kinases. Site-directed mutational analysis of the consensus N-glycosylation sites of the DDRs revealed that mutations of asparagine 213 of DDR2 and asparagine 211 of DDR1, a conserved N-glycosylation site among vertebrate DDRs, inhibited the generation of the high-molecular-mass isoform. Taken together, these results suggest a mechanism to control the activity of the DDR family by regulating its cell-surface expression. Due to low stability, the steady-state population of functional DDR proteins in the cell surface depends sensitively on its maturation process via post-translational N-glycosylation, which is controlled by the glucose supply and the presence of a conserved N-glycosylation site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Cell Line
  • Collagen / metabolism
  • Conserved Sequence*
  • Discoidin Domain Receptors
  • Endoplasmic Reticulum / metabolism
  • Extracellular Matrix / metabolism
  • Glucose / pharmacology*
  • Glycosylation / drug effects*
  • Humans
  • Male
  • Molecular Weight
  • Mutagenesis, Site-Directed
  • Phosphorylation / drug effects
  • Polysaccharides / biosynthesis
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Processing, Post-Translational / drug effects*
  • Protein Stability / drug effects
  • Protein Transport / drug effects
  • Rats
  • Receptor Protein-Tyrosine Kinases / chemistry*
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism*
  • Receptors, Mitogen / chemistry*
  • Receptors, Mitogen / genetics
  • Receptors, Mitogen / metabolism*

Substances

  • Polysaccharides
  • Protein Isoforms
  • Receptors, Mitogen
  • Collagen
  • Discoidin Domain Receptors
  • Receptor Protein-Tyrosine Kinases
  • Glucose