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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

The Machado-Joseph disease-associated expanded form of ataxin-3: Overexpression, purification, and preliminary biophysical and structural characterization

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Contessotto, Miriam G. G. [1] ; Rosselli-Murai, Luciana K. [2] ; Garcia, Maria Cristina C. [1] ; Oliveira, Cristiano L. P. [3, 4] ; Torriani, Iris L. [4, 5] ; Lopes-Cendes, Iscia [1, 6] ; Murai, Marcelo J. [1, 2]
Total Authors: 7
[1] Univ Estadual Campinas, Sch Med Sci, Dept Med Genet, Campinas, SP - Brazil
[2] Univ Michigan, Dept Pharmacol, Med Sch, Ann Arbor, MI 48109 - USA
[3] Univ Sao Paulo, Inst Phys, Sao Paulo, SP - Brazil
[4] Univ Estadual Campinas, Gleb Wataghin Inst Phys, Dept Condensed Matter Phys, Campinas, SP - Brazil
[5] Brazilian Biosci Natl Lab, Ctr Res Energy & Mat, Campinas, SP - Brazil
[6] Brazilian Inst Neurosci & Neurotechnol, Campinas, SP - Brazil
Total Affiliations: 6
Document type: Journal article
Source: Protein Expression and Purification; v. 152, p. 40-45, DEC 2018.
Web of Science Citations: 1

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. (AU)

FAPESP's process: 13/07559-3 - BRAINN - The Brazilian Institute of Neuroscience and Neurotechnology
Grantee:Fernando Cendes
Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC