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  A nanobody-based fluorescent reporter reveals human α-synuclein in the cell cytosol

Gerdes, C., Waal, N., Offner, T., Fornasiero, E. F., Wender, N., Verbarg, H., et al. (2020). A nanobody-based fluorescent reporter reveals human α-synuclein in the cell cytosol. Nature Communications, 11: 2729. doi:10.1038/s41467-020-16575-0.

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 Creators:
Gerdes, C., Author
Waal, N., Author
Offner, T., Author
Fornasiero, E. F., Author
Wender, N., Author
Verbarg, H., Author
Manzini, I., Author
Trenkwalder, C., Author
Mollenhauer, B., Author
Strohäker, T., Author
Zweckstetter, M.1, Author              
Becker, S.2, Author              
Rizzoli, S. O., Author
Basmanav, F. B., Author
Opazo, F., Author
Affiliations:
1Research Group of Protein Structure Determination using NMR, MPI for biophysical chemistry, Max Planck Society, ou_578571              
2Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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Free keywords: Biochemistry, Biomarkers, Diseases, Molecular imaging, Neuroscience
 Abstract: Aggregation and spreading of α-Synuclein (αSyn) are hallmarks of several neurodegenerative diseases, thus monitoring human αSyn (hαSyn) in animal models or cell cultures is vital for the field. However, the detection of native hαSyn in such systems is challenging. We show that the nanobody NbSyn87, previously-described to bind hαSyn, also shows cross-reactivity for the proteasomal subunit Rpn10. As such, when the NbSyn87 is expressed in the absence of hαSyn, it is continuously degraded by the proteasome, while it is stabilized when it binds to hαSyn. Here, we exploit this feature to design a new Fluorescent Reporter for hαSyn (FluoReSyn) by fusing NbSyn87 to fluorescent proteins, which results in fluorescence signal fluctuations depending on the presence and amounts of intracellular hαSyn. We characterize this biosensor in cells and tissues to finally reveal the presence of transmittable αSyn in human cerebrospinal fluid, demonstrating the potential of FluoReSyn for clinical research and diagnostics.

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Language(s): eng - English
 Dates: 2020-06-01
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41467-020-16575-0
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Title: Nature Communications
Source Genre: Journal
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Pages: - Volume / Issue: 11 Sequence Number: 2729 Start / End Page: - Identifier: -