The cell adhesion domain of type XVII collagen promotes integrin-mediated cell spreading by a novel mechanism
| Autor: | Jarmo Käpylä, Tiina Viitasalo, Jyrki Heino, Johanna Jokinen, Leena Bruckner-Tuderman, Petri Nykvist, Kaisa Tasanen |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Keratinocytes
Integrins DNA Complementary Dystonin Integrin Amino Acid Motifs Nerve Tissue Proteins CHO Cells Biochemistry Autoantigens Collagen receptor Cell Line Cell Movement Cricetinae Cell Adhesion Tumor Cells Cultured Animals Humans Cloning Molecular Cell adhesion Molecular Biology Integrin binding biology Dose-Response Relationship Drug Reverse Transcriptase Polymerase Chain Reaction Hemidesmosome Cell Biology Non-Fibrillar Collagens Molecular biology Recombinant Proteins Protein Structure Tertiary Fibronectin HaCaT Cytoskeletal Proteins Ectodomain biology.protein Collagen Carrier Proteins Peptides Protein Binding |
| Zdroj: | The Journal of biological chemistry. 276(42) |
| ISSN: | 0021-9258 3093-3099 |
| Popis: | Type XVII collagen (BP180) is a keratinocyte transmembrane protein that exists as the full-length protein in hemidesmosomes and as a 120-kDa shed ectodomain in the extracellular matrix. The largest collagenous domain of type XVII collagen, COL15, has been described previously as a cell adhesion domain (Tasanen, K., Eble, J. A., Aumailley, M., Schumann, H., Baetge, J, Tu, H., Bruckner, P., and Bruckner-Tuderman, L. (2000) J. Biol. Chem. 275, 3093-3099). In the present work, the integrin binding of triple helical, human recombinant COL15 was tested. Solid phase binding assays using recombinant integrin alpha(1)I, alpha(2)I, and alpha(10)I domains and cell spreading assays with alpha(1)beta(1)- and alpha(2)beta(1)-expressing Chinese hamster ovary cells showed that, unlike other collagens, COL15 was not recognized by the collagen receptors. Denaturation of the COL15 domain increased the spreading of human HaCaT keratinocytes, which could migrate on the denatured COL15 domain as effectively as on fibronectin. Spreading of HaCaT cells on the COL15 domain was mediated by alpha(5)beta(1) and alpha(V)beta(1) integrins, and it could be blocked by RGD peptides. The collagen alpha-chains in the COL15 domain do not contain RGD motifs but, instead, contain 12 closely related KGD motifs, four in each of the three alpha-chains. Twenty-two overlapping, synthetic peptides corresponding to the entire COL15 domain were tested; three peptides, all containing the KGD motif, inhibited the spreading of HaCaT cells on denatured COL15 domain. Furthermore, this effect was lost by mutation from D to E (KGE instead of KGD). We suggest that the COL15 domain of type XVII collagen represents a specific collagenous structure, unable to interact with the cellular receptors for other collagens. After being shed from the cell surface, it may support keratinocyte spreading and migration. |
| Databáze: | OpenAIRE |
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