α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.
Détails
ID Serval
serval:BIB_7ECFB935DE20
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.
Périodique
Journal of Biological Chemistry
ISSN
1083-351X (Electronic)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
2012
Peer-reviewed
Oui
Volume
287
Numéro
19
Pages
15345-15364
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
Since the discovery and isolation of α-synuclein (α-syn) from human brains, it has been widely accepted that it exists as an intrinsically disordered monomeric protein. Two recent studies suggested that α-syn produced in Escherichia coli or isolated from mammalian cells and red blood cells exists predominantly as a tetramer that is rich in α-helical structure (Bartels, T., Choi, J. G., and Selkoe, D. J. (2011) Nature 477, 107-110; Wang, W., Perovic, I., Chittuluru, J., Kaganovich, A., Nguyen, L. T. T., Liao, J., Auclair, J. R., Johnson, D., Landeru, A., Simorellis, A. K., Ju, S., Cookson, M. R., Asturias, F. J., Agar, J. N., Webb, B. N., Kang, C., Ringe, D., Petsko, G. A., Pochapsky, T. C., and Hoang, Q. Q. (2011) Proc. Natl. Acad. Sci. 108, 17797-17802). However, it remains unknown whether or not this putative tetramer is the main physiological form of α-syn in the brain. In this study, we investigated the oligomeric state of α-syn in mouse, rat, and human brains. To assess the conformational and oligomeric state of native α-syn in complex mixtures, we generated α-syn standards of known quaternary structure and conformational properties and compared the behavior of endogenously expressed α-syn to these standards using native and denaturing gel electrophoresis techniques, size-exclusion chromatography, and an oligomer-specific ELISA. Our findings demonstrate that both human and rodent α-syn expressed in the central nervous system exist predominantly as an unfolded monomer. Similar results were observed when human α-syn was expressed in mouse and rat brains as well as mammalian cell lines (HEK293, HeLa, and SH-SY5Y). Furthermore, we show that α-syn expressed in E. coli and purified under denaturing or nondenaturing conditions, whether as a free protein or as a fusion construct with GST, is monomeric and adopts a disordered conformation after GST removal. These results do not rule out the possibility that α-syn becomes structured upon interaction with other proteins and/or biological membranes.
Mots-clé
Amino Acid Sequence, Animals, Brain/metabolism, Cell Line, Tumor, Central Nervous System/metabolism, Chromatography, Gel, Enzyme-Linked Immunosorbent Assay, Erythrocytes/metabolism, Escherichia coli/genetics, HEK293 Cells, HeLa Cells, Humans, Immunoblotting, Mice, Mice, Transgenic, Molecular Sequence Data, Mutation, Protein Conformation, Protein Structure, Secondary, Protein Unfolding, Rats, Rats, Sprague-Dawley, Recombinant Proteins/chemistry, Recombinant Proteins/metabolism, alpha-Synuclein/chemistry, alpha-Synuclein/genetics
Pubmed
Web of science
Open Access
Oui
Création de la notice
09/11/2014 23:37
Dernière modification de la notice
20/08/2019 15:39