Nanobodies raised against monomeric ɑ-synuclein inhibit fibril formation and destabilize toxic oligomeric species

BMC Biology

Research Article

Ji-Eun Lee¹  Steven F. Lee¹  Christopher M. Dobson¹  David Klenerman¹  Marija Iljina¹  Michele Vendruscolo¹  Tuomas P. J. Knowles¹  Francesco S. Ruggeri¹  Erwin De Genst²  Tim Guilliams³  Alexander K. Buell⁴  Mathew H. Horrocks⁵  Liu Hong⁶  Marthe H. Ludtmann⁷  Sonia Gandhi⁷  Minee L. Choi⁷  Craig D. Hughes⁸  Clare E. Bryant⁸ 

[1] Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Present address: Astra Zeneca, Innovative Medicines Discovery Sciences Unit 310, Darwin Building, Cambridge Science Park, Milton Road, CB4 0WG, Cambridge, UK;Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Present address: Healx Ltd., St John’s Innovation Centre, Cowley Road, CB4 0WS, Cambridge, UK;Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Present address: Institute of Physical Biology, University of Düsseldorf, Universitätsstr. 1, 40225, Düsseldorf, Germany;Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, CB2 1QR, Cambridge, UK;Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW, Cambridge, UK;Zhou Pei-Yuan Center for Applied Mathematics, Tsinghua University, 100084, Beijing, China;Department of Molecular Neuroscience, University College London, Institute of Neurology, Queen Square, WC1N 3BG, London, UK;Department of Veterinary Medicine, University of Cambridge, Madingley Road, CB3 0ES, Cambridge, UK;

关键词: Protein aggregation;    Amyloid toxicity;    Neurodegeneration;    Aggregation inhibitors;    Antibody;    Single-molecule fluorescence;   

DOI  :  10.1186/s12915-017-0390-6

 received in 2017-04-11, accepted in 2017-06-06,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundThe aggregation of the protein ɑ-synuclein (ɑS) underlies a range of increasingly common neurodegenerative disorders including Parkinson’s disease. One widely explored therapeutic strategy for these conditions is the use of antibodies to target aggregated ɑS, although a detailed molecular-level mechanism of the action of such species remains elusive. Here, we characterize ɑS aggregation in vitro in the presence of two ɑS-specific single-domain antibodies (nanobodies), NbSyn2 and NbSyn87, which bind to the highly accessible C-terminal region of ɑS.ResultsWe show that both nanobodies inhibit the formation of ɑS fibrils. Furthermore, using single-molecule fluorescence techniques, we demonstrate that nanobody binding promotes a rapid conformational conversion from more stable oligomers to less stable oligomers of ɑS, leading to a dramatic reduction in oligomer-induced cellular toxicity.ConclusionsThe results indicate a novel mechanism by which diseases associated with protein aggregation can be inhibited, and suggest that NbSyn2 and NbSyn87 could have significant therapeutic potential.

【 授权许可】

CC BY   
© Klenerman et al. 2017

【 图 表 】

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