| Full text | |
| Author(s): |
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
|
| Affiliation: | [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: | 2 |
| 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. (AU) | |
| FAPESP's process: | 13/07559-3 - BRAINN - The Brazilian Institute of Neuroscience and Neurotechnology |
| Grantee: | Fernando Cendes |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |