Comparative in vitro and ex vivo activities of selected inhibitors of transthyretin aggregation: relevance in drug design
| Autor: | Gregorio Valencia, Gemma Arsequell, Maria João Saraiva, Maria Rosário Almeida, Isabel Cardoso, Nelson Ferreira |
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| Rok vydání: | 2007 |
| Předmět: |
Amyloid
endocrine system Protein Folding Gene Expression Plasma protein binding Transthyretin Biochemistry Mass Spectrometry Aggregation 03 medical and health sciences 0302 clinical medicine Microscopy Electron Transmission Cell Line Tumor Protein purification Animals Humans Prealbumin Molecular Biology 030304 developmental biology 0303 health sciences biology Chemistry In vitro toxicology nutritional and metabolic diseases Fibrillogenesis Cell Biology Molecular biology In vitro 3. Good health Rats Drug Design Mutation biology.protein Protein folding Anti-amyloidogenic drug Iododiflunisal 030217 neurology & neurosurgery Ex vivo Protein Binding Research Article |
| Zdroj: | Cardoso, I, Almeida, M R, Ferreira, N, Arsequell, G, Valencia, G & Saraiva, M J 2007, ' Comparative in vitro and ex vivo activities of selected inhibitors of transthyretin aggregation : Relevance in drug design ', Biochemical Journal, vol. 408, no. 1, pp. 131-138 . https://doi.org/10.1042/BJ20070689 Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1470-8728 |
| DOI: | 10.1042/BJ20070689 |
| Popis: | 8 pages, 6 figures.-- PMID: 17683281 [PubMed].-- PMCID: PMC2049069.-- Final full-text version available Open Access via PubMed Central at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17683281 Destabilization of the tetrameric fold of TTR (transthyretin) is important for aggregation of the protein which culminates in amyloid fibril formation. Many TTR mutations interfere with tetramer stability, increasing the amyloidogenic potential of the protein. The vast majority of proposed TTR fibrillogenesis inhibitors are based on in vitro assays with isolated protein, limiting their future use in clinical assays. In the present study we investigated TTR fibrillogenesis inhibitors using a cellular system that produces TTR intermediates/aggregates in the medium. Plasmids carrying wild-type TTR, V30M or L55P cDNA were transfected into a rat Schwannoma cell line and TTR aggregates were investigated in the medium using a dot-blot filter assay followed by immunodetection. Results showed that, in 24 h, TTR L55P forms aggregates in the medium, whereas, up to 72 h, wildtype TTR and V30M do not. A series of 12 different compounds, described in the literature as in vitro TTR fibrillogenesis inhibitors, were tested for their ability to inhibit L55P aggregate formation; in this system, 2-[(3,5-dichlorophenyl) amino] benzoic acid, benzoxazole, 4-(3,5-difluorophenyl) benzoic acid and triiodophenol were the most effective inhibitors, as compared with the reference iododiflunisal, previously shown by ex vivo and in vitro procedures to stabilize TTR and inhibit fibrillogenesis. Among these drugs, 2-[(3,5-dichlorophenyl) amino] benzoic acid and tri-iodophenol stabilized TTR from heterozygotic carriers of V30M in the same ex vivo conditions as those used previously for iododiflunisal. The novel cellular-based test herein proposed for TTR fibrillogenesis inhibitor screens avoids not only lengthy and cumbersome large-scale protein isolation steps but also artefacts associated with most current in vitro first-line screening methods, such as those associated with acidic conditions and the absence of serum proteins. We thank Paul Moreira for his assistance in the production of recombinant TTRs. This work was supported by the Portuguese Foundation for Science and Technology (FCT) through a fellowship to I.C. (SFRH/BPD/9416/2002), POCTI (Programa Operacional Ciência, Tecnologia, Inovaçao) and POCI (Programa Operacional Ciência e Inovaçao) grants, and by the Gulbenkian Foundation. |
| Databáze: | OpenAIRE |
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