The Machado-Joseph disease-associated expanded form of ataxin-3 : overexpression, purification, and preliminary biophysical and structural characterization
| العنوان: | The Machado-Joseph disease-associated expanded form of ataxin-3 : overexpression, purification, and preliminary biophysical and structural characterization |
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| المؤلفون: | Contessotto, Miriam Gisele Gasparotto, 1972, Garcia, Maria Cristina Cabral, Oliveira, Cristiano Luis Pinto de, 1975, Torriani, Iris Concepcion Linares de, 1934, Lopes-Cendes, Íscia Teresinha, 1964, Murai, Marcelo Jun, 1978 |
| المساهمون: | UNIVERSIDADE ESTADUAL DE CAMPINAS |
| المصدر: | Repositório Institucional da Unicamp Universidade Estadual de Campinas (UNICAMP) instacron:UNICAMP Repositório da Produção Científica e Intelectual da Unicamp |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | Doença de Machado-Joseph, Expanded ataxin-3, Machado-Joseph disease, Small-angle X-ray scattering, Artigo original, Ataxia, Raios X - Espalhamento a baixo ângulo |
| الوصف: | Agradecimentos: The authors wish to thank Tatiany Marques for technical assistance, Drs. Celso Benedetti, Avram Slovic and Allen Liu for their support. We acknowledge the patients for donation of biological material. This study was supported by the São Paulo Research Foundation grants 01/07542-6, 2013/07559-3 (IL-C), 01/12346-1 (MGGC), 01/10658-6 (MCCG) and the Brazilian National Research Council-CNPq (ILT). Beam time granted at the SAXS Beamline of the Brazilian National Synchrotron Laboratory is gratefully acknowledged Abstract: An expansion of the polyglutamine (polyQ) tract within the deubiquitinase ataxin-3 protein is believed to play a role in a neurodegenerative disorder. Ataxin-3 contains a Josephin catalytic domain and a polyQ tract that renders it intrinsically prone to aggregate, and thus full-length protein is difficult to characterize structurally by high-resolution methods. We established a robust protocol for expression and purification of wild-type and expanded ataxin-3, presenting 19Q and 74Q, respectively. Both proteins are monodisperse as assessed by analytical size exclusion chromatography. Initial biophysical characterization was performed, with apparent transition melting temperature of expanded ataxin-3 lower than the wild-type counterpart. We further characterize the molecular envelope of wild-type and expanded polyQ tract in ataxin-3 using small angle X-ray scattering (SAXS). Characterization of protein-protein interactions between ataxin-3 and newly identified binding partners will benefit from our protocol CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado |
| وصف الملف: | application/pdf |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=dedup_wf_001::3888209f15dcce4c49e8bbf2e03113caTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.dedup.wf.001..3888209f15dcce4c49e8bbf2e03113ca |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |