Identification of PTPRσ-interacting proteins by proximity-labelling assay

J Biochem. 2021 Mar 5;169(2):187-194. doi: 10.1093/jb/mvaa141.

Abstract

Receptor protein tyrosine phosphatases (RPTPs) are type-I transmembrane proteins and involved in various biological and pathological processes. Their functions are supposed to be exerted through tyrosine dephosphorylation of their specific substrates. However, our comprehensive understanding of specific substrates or interacting proteins for RPTPs is poor. PTPRσ belongs to class 2a RPTP family, dephosphorylates cortactin, and leads to autophagy flux disruption and axonal regeneration inhibition in response to its ligand chondroitin sulphate. Here, we applied proximity-dependent biotin identification (BioID) assay, a proximity-labelling assay, to PTPRσ and reproducibly identified the 99 candidates as interactors for PTPRσ including already-known interactors such as Liprin-α and Trio. Of note, cortactin was also listed up in our assay. Our results suggest that the BioID assay is a powerful and reliable tool to identify RPTP-interacting proteins including its specific substrate.

Keywords: PTPRσ; axon regeneration; protein–protein interaction; proteomics; proximity-labelling.

MeSH terms

  • Autophagy / physiology
  • Biotinylation / methods
  • Cell Line
  • Chondroitin Sulfates / metabolism*
  • HEK293 Cells
  • Humans
  • Phosphorylation
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Proteomics / methods
  • Receptor-Like Protein Tyrosine Phosphatases, Class 4 / genetics
  • Receptor-Like Protein Tyrosine Phosphatases, Class 4 / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*

Substances

  • Recombinant Fusion Proteins
  • Chondroitin Sulfates
  • PTPRA protein, human
  • Receptor-Like Protein Tyrosine Phosphatases, Class 4