Abstract
The microtubule-associated protein tau has been extensively studied as a culprit in Alzheimer’s disease and other neurodegenerative diseases known as tauopathies. Challenges in structurally defining tau protein emerge from its disordered nature, which makes it difficult to crystallize, and hinder efforts to interpret tau protein’s true function. The complexity of intrinsically disordered proteins (IDPs) necessitates a multifaceted approach to study their interactions including multiple spectroscopic methods that can report on local protein environment and structure at individual residue positions. We and others have shown that in addition to binding to microtubules, tau binds to lipid membranes. Tau-membrane interactions may be relevant both to normal tau function and to tau aggregation and pathology. Here we describe the use of fluorescence spectroscopy as a probe of protein-membrane interactions to determine whether there is an interaction, which residues participate, and the extent/nature of the interface between the protein and the membrane. We provide a protocol for how the membrane interactions of tau protein, as an example, can be probed by fluorescence spectroscopy, including details of how the samples should be prepared and guidelines on how to interpret the results.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mandelkow E-M, Mandelkow E (2012) Biochemistry and cell biology of tau protein in neurofibrillary degeneration. Cold Spring Harb Perspect Med 2:a006247
Wang Y, Mandelkow E (2016) Tau in physiology and pathology. Nat Rev Neurosci 17:22–35
Devred F, Barbier P, Douillard S et al (2004) Tau induces ring and microtubule formation from αβ-tubulin dimers under nonassembly conditions. Biochemistry 43:10520–10531
Förstl H, Kurz A (1999) Clinical features of Alzheimer’s disease. Eur Arch Psychiatry Clin Neurosci 249:288–290
Brandt R, Léger J, Lee G (1995) Interaction of tau with the neural plasma membrane mediated by tau’s amino-terminal projection domain. J Cell Biol 131:1327–1340
Shea TB (1997) Phospholipids alter tau conformation, phosphorylation, proteolysis, and association with microtubules: implication for tau function under normal and degenerative conditions. J Neurosci Res 50:114–122
Georgieva ER, Xiao S, Borbat PP et al (2014) Tau binds to lipid membrane surfaces via short amphipathic helices located in its microtubule-binding repeats. Biophys J 107:1441–1452
Barré P, Eliezer D (2006) Folding of the repeat domain of tau upon binding to lipid surfaces. J Mol Biol 362:312–326
Barré P, Eliezer D (2013) Structural transitions in tau k18 on micelle binding suggest a hierarchy in the efficacy of individual microtubule-binding repeats in filament nucleation. Protein Sci Publ Protein Soc 22:1037–1048
Ait-Bouziad N, Lv G, Mahul-Mellier A-L et al (2017) Discovery and characterization of stable and toxic tau/phospholipid oligomeric complexes. Nat Commun 8:1678
Ganguly P, Do TD, Larini L et al (2015) Tau assembly: the dominant role of PHF6 (VQIVYK) in microtubule binding region repeat R3. J Phys Chem B 119:4582–4593
Friedhoff P, von Bergen M, Mandelkow EM et al (2000) Structure of tau protein and assembly into paired helical filaments. Biochim Biophys Acta 1502:122–132
von Bergen M, Barghorn S, Li L et al (2001) Mutations of tau protein in Frontotemporal dementia promote aggregation of paired helical filaments by enhancing local β-structure. J Biol Chem 276:48165–48174
Li X-H, Culver JA, Rhoades E (2015) Tau binds to multiple tubulin dimers with helical structure. J Am Chem Soc 137:9218–9221
Eliezer D (2012) Distance information for disordered proteins from NMR and ESR measurements using paramagnetic spin labels. Methods Mol Biol (Clifton, NJ) 895:127–138
Snead D, Wragg RT, Dittman JS et al (2014) Membrane curvature sensing by the C-terminal domain of complexin. Nat Commun 5:4955
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Acosta, D., Das, T., Eliezer, D. (2020). Probing IDP Interactions with Membranes by Fluorescence Spectroscopy. In: Kragelund, B.B., Skriver, K. (eds) Intrinsically Disordered Proteins. Methods in Molecular Biology, vol 2141. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0524-0_28
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0524-0_28
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0523-3
Online ISBN: 978-1-0716-0524-0
eBook Packages: Springer Protocols