Identification of BCAP-L as a negative regulator of the TLR signaling-induced production of IL-6 and IL-10 in macrophages by tyrosine phosphoproteomics

Takayuki Matsumura, Masaaki Oyama, Hiroko Kozuka-Hata, Kosuke Ishikawa, Takafumi Inoue, Tatsushi Muta, Kentaro Senba, Jun ichiro Inoue

    Research output: Contribution to journalArticle

    13 Citations (Scopus)

    Abstract

    Toll-like receptor (TLR) signaling in macrophages is essential for anti-pathogen responses such as cytokine production and antigen presentation. Although numerous reports suggest that protein tyrosine kinases (PTKs) are involved in cytokine induction in response to lipopolysaccharides (LPS; TLR4 ligand) in macrophages, the PTK-mediated signal transduction pathway has yet to be analyzed in detail. Here, we carried out a comprehensive and quantitative dynamic tyrosine phosphoproteomic analysis on the TLR4-mediated host defense system in RAW264.7 macrophages using stable isotope labeling by amino acids in cell culture (SILAC). We determined the temporal profiles of 25 proteins based on SILAC-encoded peptide(s). Of these, we focused on the tyrosine phosphorylation of B-cell adaptor for phosphatidylinositol 3-kinase (BCAP) because the function of BCAP remains unknown in TLR signaling in macrophages. Furthermore, Bcap has two distinct transcripts, a full-length (Bcap-L) and an alternatively initiated or spliced (Bcap-S) mRNA, and little is known about the differential functions of the BCAP-L and BCAP-S proteins. Our study showed, for the first time, that RNAi-mediated selective depletion of BCAP-L enhanced IL-6 and IL-10 production but not TNF-α production in TLR ligand-stimulated macrophages. We propose that BCAP-L (but not BCAP-S) is a negative regulator of the TLR-mediated host defense system in macrophages.

    Original languageEnglish
    Pages (from-to)265-270
    Number of pages6
    JournalBiochemical and Biophysical Research Communications
    Volume400
    Issue number2
    DOIs
    Publication statusPublished - 2010 Sep

    Fingerprint

    Phosphatidylinositol 3-Kinase
    Macrophages
    Toll-Like Receptors
    Interleukin-10
    Tyrosine
    Interleukin-6
    B-Lymphocytes
    Protein-Tyrosine Kinases
    Cytokines
    Ligands
    Isotope Labeling
    Signal transduction
    Phosphorylation
    Protein S
    Antigen Presentation
    Pathogens
    RNA Interference
    Cell culture
    Isotopes
    Labeling

    Keywords

    • BCAP
    • Cytokine
    • LPS
    • Macrophage
    • SILAC
    • TLR

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Cell Biology
    • Molecular Biology
    • Medicine(all)

    Cite this

    Identification of BCAP-L as a negative regulator of the TLR signaling-induced production of IL-6 and IL-10 in macrophages by tyrosine phosphoproteomics. / Matsumura, Takayuki; Oyama, Masaaki; Kozuka-Hata, Hiroko; Ishikawa, Kosuke; Inoue, Takafumi; Muta, Tatsushi; Senba, Kentaro; Inoue, Jun ichiro.

    In: Biochemical and Biophysical Research Communications, Vol. 400, No. 2, 09.2010, p. 265-270.

    Research output: Contribution to journalArticle

    Matsumura, Takayuki ; Oyama, Masaaki ; Kozuka-Hata, Hiroko ; Ishikawa, Kosuke ; Inoue, Takafumi ; Muta, Tatsushi ; Senba, Kentaro ; Inoue, Jun ichiro. / Identification of BCAP-L as a negative regulator of the TLR signaling-induced production of IL-6 and IL-10 in macrophages by tyrosine phosphoproteomics. In: Biochemical and Biophysical Research Communications. 2010 ; Vol. 400, No. 2. pp. 265-270.
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