Scraped-Wounding Causes Activation and Association of C-Src Tyrosine Kinase with Microtubules in Cultured Keratinocytes
| Autor: | Yumi Aoyama, Takahiro Yamada, Yasuo Kitajima, Hisaaki Kawakatsu, M. Koji Owada |
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| Rok vydání: | 2000 |
| Předmět: |
Keratinocytes
Physiology Immunoblotting Cell Clone (cell biology) Sulfides SRC Family Tyrosine Kinase Biology Cell Fractionation Microtubules CSK Tyrosine-Protein Kinase Cell Movement medicine Humans Enzyme Inhibitors Cell adhesion Molecular Biology Cells Cultured Wound Healing Cell migration Cell Biology General Medicine Protein-Tyrosine Kinases Cell biology Enzyme Activation src-Family Kinases medicine.anatomical_structure Microscopy Fluorescence Cinnamates Wound healing Keratinocyte Intracellular |
| Zdroj: | Cell Structure and Function. 25:351-359 |
| ISSN: | 1347-3700 0386-7196 |
| DOI: | 10.1247/csf.25.351 |
| Popis: | In order to elucidate the function of c-Src in keratinocytes, we studied the intracellular distribution of its active and inactive form in cultured normal human keratinocyte, using anti-c-Src monoclonal antibody clone 28, which recognizes the active form of c-Src (dephosphorylated at COOH-terminal residue Tyr 530), and monoclonal antibody clone 327 which recognizes both active and inactive forms. Since c-Src has been suggested to be involved in the control of cell adhesion in other cells, we produced a dynamic condition of cell migration by cutting culture cell colonies into squares to form a mesh pattern with a blade (culture wound model). Before cutting, the active form was expressed in cells located only at the periphery of colonies or isolated migrating cells, and was associated with microtubules. Wounding the colony generated a dramatic and rapid activation of c-Src in a few rows of cells along the cut edges, which were made even at the middle of colony, resulting in the association of the active form with microtubules. This increase of the active form was also detected by immunoblotting of cell extracts. These reactions were inhibited by 1 mM sodium orthovanadate, a protein-tyrosine phosphatase inhibitor. ST 638, a potent Src family tyrosine kinase inhibitor, inhibited the migration of keratinocytes in the culture wound healing model. These results suggest that wounding the culture causes activation of c-Src in keratinocytes, and thus activated c-Src may play a role in the function of microtubules during cell migration, especially at an early stage of wound healing. |
| Databáze: | OpenAIRE |
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