ThX - A next-generation probe for the early detection of amyloid aggregates

Needham, L.-M.; Weber, J.; Varela, J. A.; Fyfe, J. W. B.; Do, D. T.; Xu, C. K.; Keenlyside, B.; Cliffe, R.; Klenerman, D.; Dobson, C. M.; Hunter, C. A.; Bohndiek, S. E.; Snaddon, T. N.; Lee, S. F.

doi:10.1101/713594

bioRxiv · 10.1101/713594

ThX - A next-generation probe for the early detection of amyloid aggregates

Needham, L.-M.·Weber, J.·Varela, J. A.·Fyfe, J. W. B.·Do, D. T.·Xu, C. K.·Keenlyside, B.·Cliffe, R.·Klenerman, D.·Dobson, C. M.·Hunter, C. A.·Bohndiek, S. E.·Snaddon, T. N.·Lee, S. F.

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Abstract

Neurodegenerative diseases such as Alzheimers and Parkinsons are associated with protein misfolding and aggregation. Recent studies suggest that the small, rare and heterogeneous oligomeric species, formed early on in the aggregation process, may be a source of cytotoxicity. Thioflavin T (ThT) is currently the gold-standard fluorescent probe for the study of amyloid proteins and aggregation processes. However, the poor photophysical and binding properties of ThT impairs the study of oligomers. To overcome this challenge, we have designed Thioflavin X, (ThX), a next-generation fluorescent probe which displays superior properties; including a 5-fold increase in brightness and 7-fold increase in binding affinity to amyloidogenic proteins. As an extrinsic dye, this can be used to study unique structural amyloid features both in bulk and on a single-aggregate level. Furthermore, ThX can be used as a super-resolution imaging probe in single-molecule localisation microscopy. Finally, we demonstrate that ThX can be used to detect a distinct oligomeric species, not observed via traditional ThT imaging.

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Needham, L.-M., Weber, J., Varela, J. A., Fyfe, J. W. B., Do, D. T., Xu, C. K., Keenlyside, B., Cliffe, R., Klenerman, D., Dobson, C. M., Hunter, C. A., Bohndiek, S. E., Snaddon, T. N., Lee, S. F.. 2019-07-24. ThX - A next-generation probe for the early detection of amyloid aggregates. https://doi.org/10.1101/713594

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