Abstract
Aggregated fibrillar tau protein is a pathological hallmark of several neurodegenerative diseases. Small molecules that bind to tau fibrils may be applied for their detection and quantification. This is of great importance as they can potentially be used for earlier diagnosis of disease and disease progression. Microscale thermophoresis (MST) enables the detection of biomolecular interactions in an aqueous environment in which no immobilization of either reaction partner is required. Here, an MST assay methodology is described for the detection of the interaction between a variety of small molecules and tau fibrils. The results of this study demonstrate that MST is a practical methodology to quantify interactions between small molecules and tau fibrillar aggregates.
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References
Vogel JW, Iturria-Medina Y, Strandberg OT et al (2020) Spread of pathological tau proteins through communicating neurons in human Alzheimer’s disease. Nat Commun 11:2612
Zhao Y, Tietz O, Kuan W-L et al (2020) A fluorescent molecular imaging probe with selectivity for soluble tau aggregated protein. Chem Sci 11:4773–4778
Harada R, Okamura N, Furumoto S, Yanai K (2018) Imaging protein misfolding in the brain using β-sheet ligands. Front Neurosci 12:585
Jerabek-Willemsen M, Wienken CJ, Braun D et al (2011) Molecular interaction studies using microscale thermophoresis. Assay Drug Dev Technol 9:342–353
Rainard JM, Pandarakalam GC, McElroy SP (2018) Using microscale thermophoresis to characterize hits from high-throughput screening: a European lead factory perspective. SLAS Discov Adv Life Sci R D 23:225–241
Jerabek-Willemsen M, André T, Wanner R et al (2014) MicroScale thermophoresis: interaction analysis and beyond. J Mol Struct 1077:101–113
Wolff M, Mittag JJ, Herling TW et al (2016) Quantitative thermophoretic study of disease-related protein aggregates. Sci Rep 6:22829
Duhr S, Braun D (2006) Why molecules move along a temperature gradient. Proc Natl Acad Sci 103:19678–19682
Seidel SAI, Dijkman PM, Lea WA et al (2013) Microscale thermophoresis quantifies biomolecular interactions under previously challenging conditions. Methods 59:301–315
Nasreddine R, Nehmé R (2021) Microscale thermophoresis for studying protein-small molecule affinity: application to hyaluronidase. Microchem J 170:106763
User Manual for the Monolith NT.115 (2017) http://www.helsinki.fi/biosciences/corefacilities/microscalethermophoresis/Manual_NT115. pdf
Fisher E, Zhao Y, Richardson R et al (2017) Detection and characterization of small molecule interactions with Fibrillar protein aggregates using microscale thermophoresis. ACS Chem Neurosci 8:2088–2095
Acknowledgments
The authors thank Prof. Alexander K. Buell for his great help with AFM imaging tau fibrils, Dr. Robert Richardson for his suggestion on preparation tau samples for MST experiments, Katalin Solti for supporting some of the MST experiments, and Prof. Finian Leeper, Mr. Ryan Bentley, and Mr. William Crone for helpful discussions. The authors thank National Institute of Health Research (NIHR, RG64473) Cambridge Biomedical Unit in Dementia, and Hungarian Brain Research Program (2017-1.2.1-NKP-2017-00002), Engineering and Physical Sciences Research Council (EP/R005397/1.) for funding.
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Zhao, Y., Mikes, A., Tóth, G., Aigbirhio, F. (2024). Detection of Small-Molecule Interactions with Fibrillar Tau Protein Aggregates Using Microscale Thermophoresis. In: Smet-Nocca, C. (eds) Tau Protein. Methods in Molecular Biology, vol 2754. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3629-9_12
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DOI: https://doi.org/10.1007/978-1-0716-3629-9_12
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-3629-9
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