Abstract
Integrins are adhesion receptors that mediate many intercellular and cell-extracellular matrix interactions with relevance in physiology and pathology. Unlike other cellular receptors, integrins critically require activation for ligand binding. Through interaction in cis with other molecules and the formation of tetraspanin-enriched membrane microdomains (TEMs), the tetraspanin CD9 regulates integrin activity and avidity. Here we present three techniques used to study CD9-integrin interactions and integrin activation.
Keywords:
CD9; Coimmunoprecipitation; Integrin; Proximity-ligation assay (PLA); Pull-down assay; Tetraspanin.
MeSH terms
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Animals
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B-Lymphocytes / cytology
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B-Lymphocytes / drug effects
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B-Lymphocytes / metabolism
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Cell Adhesion / drug effects*
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Cell Line
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Cross-Linking Reagents / chemistry
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Gene Expression
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Humans
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Immunoprecipitation / methods*
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Jurkat Cells
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Leukocytes, Mononuclear / cytology
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Leukocytes, Mononuclear / drug effects
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Leukocytes, Mononuclear / metabolism
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Lymphocyte Function-Associated Antigen-1 / genetics
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Lymphocyte Function-Associated Antigen-1 / metabolism*
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Primary Cell Culture
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Protein Binding
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Succinimides / chemistry
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THP-1 Cells
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Tetradecanoylphorbol Acetate / pharmacology
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Tetraspanin 28 / genetics
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Tetraspanin 28 / metabolism*
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Tetraspanin 29 / genetics
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Tetraspanin 29 / metabolism*
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Tetraspanin 30 / genetics
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Tetraspanin 30 / metabolism*
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U937 Cells
Substances
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CD63 protein, human
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CD81 protein, human
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CD9 protein, human
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Cross-Linking Reagents
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Lymphocyte Function-Associated Antigen-1
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Recombinant Proteins
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Succinimides
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Tetraspanin 28
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Tetraspanin 29
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Tetraspanin 30
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3,3'-dithiobis(sulfosuccinimidyl propionate)
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Tetradecanoylphorbol Acetate