Proteomic identification of common SCF ubiquitin ligase FBXO6-interacting glycoproteins in three kinds of cells

J Proteome Res. 2012 Mar 2;11(3):1773-81. doi: 10.1021/pr2010204. Epub 2012 Feb 10.


FBOX6 ubiquitin ligase complex is involved in the endoplasmic reticulum-associated degradation pathway by mediating the ubiquitination of glycoproteins. FBXO6 interacts with the chitobiose in unfolded N-glycoprotein, pointing glycoproteins toward E2 for ubiquitination. Although the glycoprotein-recognizing mechanism of FBXO6 is well documented, its bona fide interacting glycoproteins are largely unknown. Here we utilized a protein purification approach combined with LC-MS to systematically identify the FBXO6-interacting glycoproteins. Following identification of 39 proteins that specifically interact with FBXO6 in all three different cell lines, 293T, HeLa and Jurkat cells, we compared the protein complex organization between wild-type FBXO6 and its mutant, which fails to recognize glycoproteins. Combining these databases, 29 highly confident glycoproteins that interact with FBXO6 in an N-glycan dependent manner are identified. Our data provide valuable information for the discovery of the interacting glycoproteins of FBXO6 and also demonstrate the potential of these approaches as general platforms for the global discovery of interacting glycoproteins of other FBAs (F-box associated regions) containing F-box proteins.

MeSH terms

  • Cullin Proteins / metabolism
  • Glycoproteins / isolation & purification
  • Glycoproteins / metabolism*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Jurkat Cells
  • Membrane Glycoproteins / metabolism
  • Oxidoreductases / metabolism
  • Protein Binding
  • Protein Interaction Mapping
  • Protein Interaction Maps
  • Proteomics
  • SKP Cullin F-Box Protein Ligases / metabolism*


  • Cullin 1
  • Cullin Proteins
  • Glycoproteins
  • Membrane Glycoproteins
  • ERO1A protein, human
  • Oxidoreductases
  • FBXO6 protein, human
  • SKP Cullin F-Box Protein Ligases