α-Synuclein in Central Nervous System and from Erythrocytes, Mammalian Cells, and Escherichia coli Exists Predominantly as Disordered Monomer

被引：440

作者：

Fauvet, Bruno ^{[1
]}

Mbefo, Martial K. ^{[1
]}

Fares, Mohamed-Bilal ^{[1
]}

Desobry, Carole ^{[1
]}

Michael, Sarah ^{[2
]}

Ardah, Mustafa T. ^{[3
]}

Tsika, Elpida ^{[4
]}

Coune, Philippe ^{[5
]}

Prudent, Michel ^{[6
]}

Lion, Niels ^{[6
]}

Eliezer, David ^{[7
,8
]}

Moore, Darren J. ^{[4
]}

Schneider, Bernard ^{[5
]}

Aebischer, Patrick ^{[5
]}

El-Agnaf, Omar M. ^{[3
]}

Masliah, Eliezer ^{[2
]}

Lashuel, Hilal A. ^{[1
]}

机构：

[1] Ecole Polytech Fed Lausanne, Lab Mol & Chem Biol Neurodegenerat, Sch Life Sci, Brain Mind Inst,Stn 19, CH-1015 Lausanne, Switzerland

[2] Univ Calif San Diego, Sch Med, Dept Neurosci, La Jolla, CA 92093 USA

[3] United Arab Emirates Univ, Fac Med & Hlth Sci, Dept Biochem, Al Ain 15551, U Arab Emirates

[4] Ecole Polytech Fed Lausanne, Lab Mol Neurodegenerat Res, CH-1015 Lausanne, Switzerland

[5] Ecole Polytech Fed Lausanne, Neurodegenerat Dis Lab, CH-1015 Lausanne, Switzerland

[6] Serv Reg Vaudois Transfus Sanguine, CH-1066 Epalinges, Switzerland

[7] Weill Cornell Med Coll, Dept Biochem, New York, NY 10065 USA

[8] Weill Cornell Med Coll, Program Struct Biol, New York, NY 10065 USA

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2012年 / 287卷 / 19期

基金：

瑞士国家科学基金会; 美国国家卫生研究院;

关键词：

GLUTATHIONE-S-TRANSFERASE; FAMILIAL PARKINSONS-DISEASE; IN-VITRO; 3-DIMENSIONAL STRUCTURE; SELF-OLIGOMERIZATION; ALZHEIMERS-DISEASE; RESIDUAL STRUCTURE; TRANSGENIC MICE; EXTENDED-HELIX; CROSS-LINKING;

D O I：

10.1074/jbc.M111.318949

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Since the discovery and isolation of alpha-synuclein (alpha-syn) from human brains, it has been widely accepted that it exists as an intrinsically disordered monomeric protein. Two recent studies suggested that alpha-syn produced in Escherichia coli or isolated from mammalian cells and red blood cells exists predominantly as a tetramer that is rich in alpha-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 alpha-syn in the brain. In this study, we investigated the oligomeric state of alpha-syn in mouse, rat, and human brains. To assess the conformational and oligomeric state of native alpha-syn in complex mixtures, we generated alpha-syn standards of known quaternary structure and conformational properties and compared the behavior of endogenously expressed alpha-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 alpha-syn expressed in the central nervous system exist predominantly as an unfolded monomer. Similar results were observed when human alpha-syn was expressed in mouse and rat brains as well as mammalian cell lines (HEK293, HeLa, and SH-SY5Y). Furthermore, we show that alpha-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 alpha-syn becomes structured upon interaction with other proteins and/or biological membranes.

引用

页码：15345 / 15364

页数：20

共 73 条

[1] Phosphorylation of Ser-129 is the dominant pathological modification of α-synuclein in familial and sporadic Lewy body disease [J].