The C19S Substitution Enhances the Stability of Hepcidin While Conserving Its Biological Activity
| Autor: | Zsuzsanna Fekete, Edit Varga, Kitti Tamási, Katalin Sipos, Edina Pandur, László Grama |
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| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular inorganic chemicals 0301 basic medicine congenital hereditary and neonatal diseases and abnormalities media_common.quotation_subject Ferroportin Mutant Bioengineering digestive system Biochemistry Cell Line Analytical Chemistry Serine 03 medical and health sciences 0302 clinical medicine Hepcidins Western blot Hepcidin hemic and lymphatic diseases medicine Humans Cysteine Receptor Internalization media_common biology medicine.diagnostic_test Protein Stability Chemistry Organic Chemistry nutritional and metabolic diseases Biological activity 030104 developmental biology Amino Acid Substitution 030220 oncology & carcinogenesis biology.protein |
| Zdroj: | The Protein Journal. 37:113-121 |
| ISSN: | 1875-8355 1572-3887 |
| DOI: | 10.1007/s10930-018-9759-9 |
| Popis: | Hepcidin, the key hormone of iron homeostasis is responsible for lowering the serum iron level through its interaction with iron exporter ferroportin. Thus, hepcidin agonists provide a promising opportunity in the treatment of iron disorders caused by lacking or decreased hepcidin expression. We investigated the importance of each of the eight highly conserved cysteines for the biological activity of hepcidin. Eight cysteine mutants were created with site directed mutagenesis. The binding ability of these hepcidin mutants to the hepcidin receptor ferroportin was determined using bacterial two-hybrid system and WRL68 human hepatic cells. The biological activity of hepcidin mutants was determined by western blot analysis of ferroportin internalization and ferroportin ubiquitination. To investigate the effect of mutant hepcidins on the iron metabolism of the WRL68 cells, total intracellular iron content was measured with a colorimetric assay. The stability of M6 hepcidin mutant was determined using ELISA technique. Our data revealed that serine substitution of the sixth cysteine (M6) yielded a biologically active but significantly more stable peptide than the original hormone. This result may provide a promising hepcidin agonist worth testing in animal models. |
| Databáze: | OpenAIRE |
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