The intracellular signal transduction mechanism of interleukin 5 in eosinophils: The involvement of lyn tyrosine kinase and the Ras-Raf-1-MEK-microtubule-associated protein kinase pathway

Konrad Pazdrak, Douglas Schreiber, Patricia Forsythe, Louis Justement, Rafeul Alam

    Research output: Contribution to journalArticle

    148 Citations (Scopus)

    Abstract

    Interleukin 5 (IL-5) regulates the growth and function of eosinophils. The objective of this study was to investigate the intracellular signal transduction mechanism of IL-5 in eosinophils. Purified eosinophils were stimulated with IL-5, and the involvement of various kinases was investigated by immunoblotting, immune complex kinase assay, and in situ denatured/renatured kinase assay. We found that IL-5 induced tyrosine phosphorylation and activation of a number of kinases. Two species of lyn kinases (53 and 56 kD) were present in eosinophils. Both forms were Tyr-phosphorylated and activated rapidly within 1 min. Further, lyn kinase was physically associated with the IL-5β receptor in eosinophils. Ras was studied by immunoprecipitation followed by thin-layer chromatography. Ras bound higher quantities of [α-32P]guanosine 5′-triphosphate upon stimulation with IL-5. Raf-1 kinase showed increased Tyr phosphorylation on immunoblotting and increased activity in the immune complex kinase assay. Two species of MEK (MAP or Erk kinase) (41 and 45 kD) were identified in eosinophils, which underwent autophosphorylation upon stimulation. Microtubule-associated protein (MAP) kinase (p44) was Tyr-phosphorylated on immunoblotting and had increased activity in the immune-complex kinase assay. MAP kinases were also studied after metabolic radiolabeling of the cells with [32P] orthophosphates. IL-5 stimulated phosphorylation of MAP kinases in situ. Thus, we have delineated major components of an important signaling pathway in eosinophils. We believe that one of the signals generated by IL-5 receptor activation is propagated through the lyn-Ras-Raf-1-MEK-MAP kinase pathway.

    Original languageEnglish (US)
    Pages (from-to)1827-1834
    Number of pages8
    JournalJournal of Experimental Medicine
    Volume181
    Issue number5
    StatePublished - May 1 1995

    Fingerprint

    Proto-Oncogene Proteins c-raf
    Microtubule-Associated Proteins
    Interleukin-5
    Eosinophils
    Protein-Tyrosine Kinases
    Protein Kinases
    Signal Transduction
    Phosphotransferases
    Interleukin-5 Receptors
    Antigen-Antibody Complex
    Immunoblotting
    Phosphorylation
    Thin Layer Chromatography
    Guanosine Triphosphate
    Immunoprecipitation
    Tyrosine
    Phosphates

    ASJC Scopus subject areas

    • Immunology

    Cite this

    The intracellular signal transduction mechanism of interleukin 5 in eosinophils : The involvement of lyn tyrosine kinase and the Ras-Raf-1-MEK-microtubule-associated protein kinase pathway. / Pazdrak, Konrad; Schreiber, Douglas; Forsythe, Patricia; Justement, Louis; Alam, Rafeul.

    In: Journal of Experimental Medicine, Vol. 181, No. 5, 01.05.1995, p. 1827-1834.

    Research output: Contribution to journalArticle

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