α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.

Details

Serval ID
serval:BIB_7ECFB935DE20
Type
Article: article from journal or magazin.
Collection
Publications
Title
α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.
Journal
Journal of Biological Chemistry
Author(s)
Fauvet B., Mbefo M.K., Fares M.B., Desobry C., Michael S., Ardah M.T., Tsika E., Coune P., Prudent M., Lion N., Eliezer D., Moore D.J., Schneider B., Aebischer P., El-Agnaf O.M., Masliah E., Lashuel H.A.
ISSN
1083-351X (Electronic)
ISSN-L
0021-9258
Publication state
Published
Issued date
2012
Peer-reviewed
Oui
Volume
287
Number
19
Pages
15345-15364
Language
english
Notes
Publication types: Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
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.
Keywords
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
Yes
Create date
09/11/2014 22:37
Last modification date
20/08/2019 14:39
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