Probing Structure and Function of Alkali Sensor IRR with Monoclonal Antibodies
| Autor: | Alexander N. Orsa, Tatiana N. Erokhina, I. E. Deyev, Alexander S. Goryashchenko, O. V. Serova, Alexander G. Petrenko, A. A. Mozhaev |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Models Molecular Immunoprecipitation medicine.drug_class Protein Conformation IRR Blotting Western lcsh:QR1-502 Alkalies Monoclonal antibody Biochemistry Receptor tyrosine kinase lcsh:Microbiology Article activator 03 medical and health sciences Mice 0302 clinical medicine Protein Domains parasitic diseases medicine Animals Humans Binding site Receptor Molecular Biology Orphan receptor biology Chemistry Antibodies Monoclonal Molecular biology Immunohistochemistry Receptor Insulin inhibitor Insulin receptor alkali sensor 030104 developmental biology HEK293 Cells Ectodomain monoclonal antibody 030220 oncology & carcinogenesis biology.protein receptor tyrosine kinase population characteristics |
| Zdroj: | Biomolecules Biomolecules, Vol 10, Iss 1060, p 1060 (2020) Volume 10 Issue 7 |
| ISSN: | 2218-273X |
| Popis: | To study the structure and function of the pH-regulated receptor tyrosine kinase insulin receptor-related receptor (IRR), а member of the insulin receptor family, we obtained six mouse monoclonal antibodies against the recombinant IRR ectodomain. These antibodies were characterized in experiments with exogenously expressed full-length IRR by Western blotting, immunoprecipitation, and immunocytochemistry analyses. Utilizing a previously obtained set of IRR/IR chimeras with swapped small structural domains and point amino acid substitutions, we mapped the binding sites of the obtained antibodies in IRR. Five of them showed specific binding to different IRR domains in the extracellular region, while one failed to react with the full-length receptor. Unexpectedly, we found that 4D5 antibody can activate IRR at neutral pH, and 4C2 antibody can inhibit activation of IRR by alkali. Our study is the first description of the instruments of protein nature that can regulate activity of the orphan receptor IRR and confirms that alkali-induced activation is an intrinsic property of this receptor tyrosine kinase. |
| Databáze: | OpenAIRE |
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