IRIS Catalogo Istituzionale della Ricerca dell'Università degli Studi del Molise
IRIS è il Catalogo Istituzionale della Ricerca dell'Università degli Studi del Molise ed ha lo scopo di consentire la conservazione e la consultazione delle informazioni sulla produzione scientifica dell'Ateneo.
EnglishFerroportin (Fpn) (IREG1, SLC40A1, MTP1) is an iron transporter, and mutations in Fpn result in a genetically dominant form of iron overload disease. Previously, we demonstrated that Fpn is a multimer and that mutations in Fpn are dominant negative. Other studies have suggested that Fpn is not a multimer and that overexpression or epitope tags might affect the localization, topology, or multimerization of Fpn. We generated wildtype Fpn with 3 different epitopes, GFP, FLAG, and c-myc, and expressed these constructs in cultured cells. Co-expression of any 2 different epitope-tagged proteins in the same cell resulted in their quantitative coimmunoprecipitation. Treatment of Fpn-GFP/Fpn-FLAG–expressing cells with crosslinking reagents resulted in the crosslinking of Fpn-GFP and Fpn-FLAG. Western analysis of rat glioma C6 cells or mousebone marrow macrophages exposed to crosslinking reagentsshowedthat endogenous Fpn is a dimer. These results support the hypothesis that the dominant inheritance of Fpn–iron overload disease is due to the dominant-negative effects of mutant Fpn proteins.
| Codice identificativo ISI: | WOS:000244641100064 |
| Codice identificativo Scopus: | 2-s2.0-33847412861 |
| Handle: | http://hdl.handle.net/11695/6824 |
| Abstract: | Ferroportin (Fpn) (IREG1, SLC40A1, MTP1) is an iron transporter, and mutations in Fpn result in a genetically dominant form of iron overload disease. Previously, we demonstrated that Fpn is a multimer and that mutations in Fpn are dominant negative. Other studies have suggested that Fpn is not a multimer and that overexpression or epitope tags might affect the localization, topology, or multimerization of Fpn. We generated wildtype Fpn with 3 different epitopes, GFP, FLAG, and c-myc, and expressed these constructs in cultured cells. Co-expression of any 2 different epitope-tagged proteins in the same cell resulted in their quantitative coimmunoprecipitation. Treatment of Fpn-GFP/Fpn-FLAG–expressing cells with crosslinking reagents resulted in the crosslinking of Fpn-GFP and Fpn-FLAG. Western analysis of rat glioma C6 cells or mousebone marrow macrophages exposed to crosslinking reagentsshowedthat endogenous Fpn is a dimer. These results support the hypothesis that the dominant inheritance of Fpn–iron overload disease is due to the dominant-negative effects of mutant Fpn proteins. |
| Titolo: | Evidence for the multimeric structure of ferroportin |
| Autori: | |
| Appare nelle tipologie: | 1.1 Articolo in rivista |
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