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  • The Lyn kinase C-lobe mediates Golgi export of Lyn through conformation-dependent ACSL3 association. 20605918

    The Src-family tyrosine kinase Lyn has a role in signal transduction at the cytoplasmic face of the plasma membrane upon extracellular ligand stimulation. After synthesis in the cytoplasm, Lyn accumulates on the Golgi and is subsequently transported to the plasma membrane. However, the mechanism of Lyn trafficking remains elusive. We show here that the C-lobe of the Lyn kinase domain is associated with long-chain acyl-CoA synthetase 3 (ACSL3) on the Golgi in a manner that is dependent on Lyn conformation but is independent of its kinase activity. Formation of a closed conformation by CSK prevents Lyn from associating with ACSL3, resulting in blockade of Lyn export from the Golgi. Overexpression and knockdown of ACSL3 accelerates and blocks Golgi export of Lyn, respectively. The post-Golgi route of Lyn, triggered by ACSL3, is distinct from that of vesicular stomatitis virus glycoprotein (VSV-G) and of caveolin. Moreover, an ACSL3 mutant lacking the LR2 domain, which is required for the catalytic activity, retains the ability to associate with Lyn and accelerate Golgi export of Lyn. These results suggest that initiation of Golgi export of Lyn involves association of ACSL3 with the Lyn C-lobe, which is exposed to the molecular surface in an open conformation.
    Document Type:
    Reference
    Product Catalog Number:
    MAB1501
    Product Catalog Name:
    Anti-Actin Antibody, clone C4

  • Lyn, a Src family kinase, regulates activation of epidermal growth factor receptors in lung adenocarcinoma cells. 23866081

    Activation of receptors for growth factors on lung epithelial cells is essential for transformation into tumor cells, supporting their viability and proliferation. In most lung cancer patients, EGFR is constitutively activated without evidence of mutation. Defining mechanisms for constitutive activation of EGFR could elucidate additional targets for therapy of lung cancers.The approach was to identify lung cancer cell lines with constitutively activated EGFR and use systematic selection of inhibitors to evaluate their effects on specific EGFR phosphorylations and downstream signaling pathways. Interactions between receptors, kinases, and scaffolding proteins were investigated by co-immunoprecipitation plus Western blotting.The results revealed a dependence on Src family of tyrosine kinases for downstream signaling and cell growth. Lyn, a Src family kinase functional in normal and malignant B-lymphocytes, was a defining signal transducer required for EGFR signaling in Calu3 cell line. Src family kinase activation in turn, was dependent on PKCßII. Lyn and PKC exist in membrane complexes of RACK1 and in association with EGFR which pairs with other receptor partners. Silencing of Lyn expression with interfering siRNA decreased EGFR activation and cell viability.The importance of Src family kinases and PKCßII in the initiation of the EGFR signaling pathway in lung tumor cells was demonstrated. We conclude that phosphorylation of EGFR is mediated through PKCßII regulation of Lyn activation, and occurs in association with RACK1 and Cbp/PAG proteins. We suggest that protein complexes in cell membranes, including lipid rafts, may serve as novel targets for combination therapies with EGFR and Src Family Kinase inhibitors in lung cancer.
    Document Type:
    Reference
    Product Catalog Number:
    48-650MAG
    Product Catalog Name:
    MILLIPLEX® 8-Plex Human Src Family Kinase Kit – Phosphoprotein - Cancer Multiplex Assay

  • LY 294002

    Document Type:
    Certificate of Analysis
    Lot Number:
    2485691
    Product Catalog Number:
    19-142
    Product Catalog Name:
    LY 294002

  • LY 294002 - 3722124

    Document Type:
    Certificate of Analysis
    Lot Number:
    3722124
    Product Catalog Number:
    19-142
    Product Catalog Name:
    LY 294002

  • Lyn, active - 1660215-F

    Document Type:
    Certificate of Analysis
    Lot Number:
    1660215-F
    Product Catalog Number:
    14-510M
    Product Catalog Name:
    Lyn Protein, active, 250 µg

  • LY 294002 - 2836189

    Document Type:
    Certificate of Analysis
    Lot Number:
    2836189
    Product Catalog Number:
    19-142
    Product Catalog Name:
    LY 294002

  • Lyn modulates Claudin-2 expression and is a therapeutic target for breast cancer liver metastasis. 25823815

    Claudin-2 enhances breast cancer liver metastasis and promotes the development of colorectal cancers. The objective of our current study is to define the regulatory mechanisms controlling Claudin-2 expression in breast cancer cells. We evaluated the effect of several Src Family Kinase (SFK) inhibitors or knockdown of individual SFK members on Claudin-2 expression in breast cancer cells. We also assessed the potential effects of pan-SFK and SFK-selective inhibitors on the formation of breast cancer liver metastases. This study reveals that pan inhibition of SFK signaling pathways significantly elevated Claudin-2 expression levels in breast cancer cells. In addition, our data demonstrate that pan-SFK inhibitors can enhance breast cancer metastasis to the liver. Knockdown of individual SFK members reveals that loss of Yes or Fyn induces Claudin-2 expression; whereas, diminished Lyn levels impairs Claudin-2 expression in breast cancer cells. The Lyn-selective kinase inhibitor, Bafetinib (INNO-406), acts to reduce Claudin-2 expression and suppress breast cancer liver metastasis. Our findings may have major clinical implications and advise against the treatment of breast cancer patients with broad-acting SFK inhibitors and support the use of Lyn-specific inhibitors.
    Document Type:
    Reference
    Product Catalog Number:
    05-184
    Product Catalog Name:
    Anti-Src Antibody, clone GD11