CIN85 interacting proteins in B cells-specific role for SHIP-1

Mol Cell Proteomics. 2011 Oct;10(10):M110.006239. doi: 10.1074/mcp.M110.006239. Epub 2011 Jul 1.

Abstract

The Cbl-interacting 85-kDa protein (CIN85) plays an important role as a negative regulator of signaling pathways induced by receptor tyrosine kinases. By assembling multiprotein complexes this versatile adaptor enhances receptor tyrosine kinase-activated clathrin-mediated endocytosis and reduces phosphatidylinositol-3-kinase-induced phosphatidylinositol-3,4,5-trisphosphate production. Here we report the expression of CIN85 in primary splenic B lymphocytes and the B-lymphoma cell lines WEHI 231 and Ba/F3. Cross-linking of the B cell antigen receptor resulted in an increased association of CIN85 with the ubiquitin ligase Cbl. Through a systematic pull-down proteomics approach we identified 51 proteins that interact with CIN85 in B cells, including proteins not shown previously to be CIN85-associated. Among these proteins, the SH2-containing inositol phosphatase 1 (SHIP-1) co-precipitated with both the full-length CIN85 and each of its three SH3 domains. We also showed that this association is constitutive and depends on a region of 79 amino acids near the carboxyl terminus of SHIP-1, a region rich in potential SH3 domain binding sites. Because SHIP-1 is a major negative regulator of the phosphatidylinositol-3-kinase pathway in lymphocytes, we hypothesize that the interaction between SHIP-1 and CIN85 might synergistically facilitate the down-regulation of phosphatidylinositol-3,4,5-trisphosphate levels.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • B-Lymphocytes / metabolism
  • Binding Sites
  • Cell Line, Tumor
  • Gene Expression Regulation
  • Humans
  • Inositol Polyphosphate 5-Phosphatases
  • Jurkat Cells
  • Male
  • Mass Spectrometry
  • Mice
  • Mice, Inbred C57BL
  • Multiprotein Complexes / metabolism
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Phosphatidylinositol 3-Kinase / metabolism*
  • Phosphatidylinositol Phosphates / chemistry
  • Phosphatidylinositol Phosphates / metabolism*
  • Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism*
  • Protein Binding
  • Proteomics
  • Proto-Oncogene Proteins c-cbl / metabolism*
  • Recombinant Proteins / metabolism
  • Signal Transduction
  • Spleen / cytology
  • src Homology Domains

Substances

  • Adaptor Proteins, Signal Transducing
  • Multiprotein Complexes
  • Neoplasm Proteins
  • Nerve Tissue Proteins
  • Phosphatidylinositol Phosphates
  • Recombinant Proteins
  • Sh3kbp1 protein, mouse
  • phosphatidylinositol 3,4,5-triphosphate
  • Proto-Oncogene Proteins c-cbl
  • Phosphatidylinositol 3-Kinase
  • Phosphoric Monoester Hydrolases
  • Inositol Polyphosphate 5-Phosphatases
  • INPP5D protein, human
  • Inpp5d protein, mouse
  • Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases