Requirement of Gamma-Carboxyglutamic Acid Modification and Phosphatidylserine Binding for the Activation of Tyro3, Axl, and Mertk Receptors by Growth Arrest-Specific 6
| Autor: | Canan Kasikara, Oleta A. Sandiford, Kensaku Mizuno, Stanley G. Kimani, Pranela Rameshwar, Sushil Kumar, Raymond B. Birge, Ke Geng, Vladyslav Kholodovych, Sergei V. Kotenko |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
lcsh:Immunologic diseases. Allergy phosphatidylserine γ-carboxylation Immunology tumor exosomes Biology vitamin K 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine stomatognathic system Immunology and Allergy Receptor Efferocytosis skin and connective tissue diseases Phosphatidylserine binding Original Research Gla domain GAS6 Axl Phosphatidylserine MERTK Tyro3 Cell biology 030104 developmental biology Biochemistry chemistry 030220 oncology & carcinogenesis and Mertk receptors growth arrest-specific 6 lcsh:RC581-607 Tyrosine kinase hormones hormone substitutes and hormone antagonists |
| Zdroj: | Frontiers in Immunology Frontiers in Immunology, Vol 8 (2017) |
| ISSN: | 1664-3224 |
| Popis: | The Tyro3, Axl, and Mertk (TAM) receptors are homologous type I receptor tyrosine kinases that have critical functions in the clearance of apoptotic cells in multicellular organisms. TAMs are activated by their endogenous ligands, growth arrest-specific 6 (Gas6), and protein S (Pros1), that function as bridging molecules between externalized phosphatidylserine (PS) on apoptotic cells and the TAM ectodomains. However, the molecular mechanisms by which Gas6/Pros1 promote TAM activation remains elusive. Using TAM/IFNγR1 reporter cell lines to monitor functional TAM activity, we found that Gas6 activity was exquisitely dependent on vitamin K-mediated γ-carboxylation, whereby replacing vitamin K with anticoagulant warfarin, or by substituting glutamic acid residues involved in PS binding, completely abrogated Gas6 activity as a TAM ligand. Furthermore, using domain and point mutagenesis, Gas6 activity also required both an intact Gla domain and intact EGF-like domains, suggesting these domains function cooperatively in order to achieve TAM activation. Despite the requirement of γ-carboxylation and the functional Gla domain, non-γ-carboxylated Gas6 and Gla deletion/EGF-like domain deletion mutants still retained their ability to bind TAMs and acted as blocking decoy ligands. Finally, we found that distinct sources of PS-positive cells/vesicles (including apoptotic cells, calcium-induced stressed cells, and exosomes) bound Gas6 and acted as cell-derived or exosome-derived ligands to activate TAMs. Taken together, our findings indicate that PS is indispensable for TAM activation by Gas6, and by inference, provides new perspectives on how PS, regulates TAM receptors and efferocytosis. |
| Databáze: | OpenAIRE |
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