Abstract
Membrane interactions of proteins play a role in essential cellular processes in both physiological and disease states. The structural flexibility of intrinsically disordered proteins (IDPs) allows for interactions with multiple partners, including membranes. However, determining conformational states of IDPs when interacting with membranes can be challenging. Here we describe the use of nuclear magnetic resonance (NMR), including dark-state exchange saturation transfer (DEST), to probe IDP-membrane interactions in order to determine whether there is an interaction, which residues participate, and the extent/nature of the interaction between the protein and the membrane. Using α-synuclein and tau as typical examples, we provide protocols for how the membrane interactions of IDPs can be probed, including details of how the samples should be prepared and guidelines on how to interpret the results.
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References
Duplâtre G, Ferreira Marques MF, Da Graça Miguel M (1996) Size of sodium dodecyl sulfate micelles in aqueous solutions as studied by positron annihilation lifetime spectroscopy. J Phys Chem 100:16608–16612
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 22:1037–1048
Bussell R, Eliezer D (2003) A structural and functional role for 11-mer repeats in α-synuclein and other exchangeable lipid binding proteins. J Mol Biol 329(4):763–778
Snead D, Eliezer D (2018) Spectroscopic characterization of structure–function relationships in the intrinsically disordered protein complexin. In: Methods in enzymology. Elsevier, Amsterdam
Fawzi NL, Ying J, Torchia DA et al (2012) Probing exchange kinetics and atomic resolution dynamics in high-molecular-weight complexes using dark-state exchange saturation transfer NMR spectroscopy. Nat Protoc 7:1523–1533
Georgieva ER, Ramlall TF, Borbat PP et al (2010) The lipid-binding domain of wild type and mutant alpha-synuclein: compactness and interconversion between the broken and extended helix forms. J Biol Chem 285:28261–28274
Georgieva ER, Ramlall TF, Borbat PP et al (2008) Membrane-bound alpha-synuclein forms an extended helix: long-distance pulsed ESR measurements using vesicles, bicelles, and rodlike micelles. J Am Chem Soc 130:12856–12857
Jao CC, Hegde BG, Chen J et al (2008) Structure of membrane-bound alpha-synuclein from site-directed spin labeling and computational refinement. Proc Natl Acad Sci U S A 105:19666–19671
Eliezer D (2012) Distance information for disordered proteins from NMR and ESR measurements using paramagnetic spin labels. Methods Mol Biol 895:127–138
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
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(5):1327–1340
Gauthier-Kemper A, Weissmann C, Golovyashkina N et al (2011) The frontotemporal dementia mutation R406W blocks tau’s interaction with the membrane in an annexin A2-dependent manner. J Cell Biol 192(4):647–661
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
Gray EG, Paula-Barbosa M, Roher A (1987) Alzheimer’s disease: paired helical filaments and cytomembranes. Neuropathol Appl Neurobiol 13:91–110
Barré P, Eliezer D (2006) Folding of the repeat domain of tau upon binding to lipid surfaces. J Mol Biol 362:312–326
Emanuele M, Chieregatti E (2015) Mechanisms of alpha-synuclein action on neurotransmission: cell-autonomous and non-cell autonomous role. Biomolecules 5(2):865–892
Das T, Eliezer D (2019) Membrane interactions of intrinsically disordered proteins: the example of alpha-synuclein. Biochim Biophys Acta Proteins Proteom 1867:879–889
Gonzalez-Horta A, Gonzalez Hernandez B, Chavez-Montes A (2013) Fluorescence as a tool to study lipid-protein interactions: the case of α-Synuclein. Open J Biophys 3:112–119
Das T, Eliezer D (2019) Probing structural changes in alpha-synuclein by nuclear magnetic resonance spectroscopy. Methods Mol Biol 1948:157–181
Fawzi NL, Ying J, Ghirlando R et al (2011) Atomic resolution dynamics on the surface of amyloid β protofibrils probed by solution NMR. Nature 480:268–272
Delaglio F, Grzesiek S, Vuister GW et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293
Eliezer D, Barré P, Kobaslija M et al (2005) Residual structure in the repeat domain of tau: Echoes of microtubule binding and paired helical filament formation. Biochemistry 44(3):1026–1036
Harbison NW, Bhattacharya S, Eliezer D (2012) Assigning backbone NMR resonances for full length tau isoforms: efficient compromise between manual assignments and reduced dimensionality. PLoS One 7:e34679
Anthis NJ, Clore GM (2015) Visualizing transient dark states by NMR spectroscopy. Q Rev Biophys 48:35–116
Maltsev AS, Chen J, Levine RL et al (2013) Site-specific interaction between α-synuclein and membranes probed by NMR-observed methionine oxidation rates. J Am Chem Soc 135:2943–2946
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Das, T., Acosta, D., Eliezer, D. (2020). Interactions of IDPs with Membranes Using Dark-State Exchange NMR 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_30
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DOI: https://doi.org/10.1007/978-1-0716-0524-0_30
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