Depicting Conformational Ensembles of α-Synuclein by Single Molecule Force Spectroscopy and Native Mass Spectroscopy
Abstract
:1. Introduction
2. Results
2.1. Single Molecule Force Spectroscopy (SMSF)
2.2. Effect of DA and EGCG on the Conformational Ensemble
2.3. Comparison to CD and FTIR
3. Discussion
4. Materials and Methods
4.1. Cloning, Expression, and Purification of the (I27)4_AS_(I27)4 Polyprotein
4.2. AFM—Single Molecule Force Spectroscopy
4.3. AFM Data Analysis
4.4. Native-MS Experiments
4.5. CD and FTIR Experiments
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AS | α-synuclein |
C | compact structure detected in native MS |
CD | circular dichroism |
CSDs | charge state distributions |
DA | dopamine |
EGCG | epigallocatechin-3-gallate |
ESI-MS | electrospray ionization mass spectrometry |
F | unfolding force |
FTIR | Fourier-transform infrared spectroscopy |
I1, I2 | Intermediate 1 and 2 detected in native MS |
I27 | 27th titin immunoglobulin-like domain |
IDP | intrinsically disordered protein |
IDPs | intrinsically disordered proteins |
LC | contour length |
LP | persistence length |
native MS | native mass spectrometry |
NMR | nuclear magnetic resonance |
RC | random coil |
SAXS-EOM | small-angle X-ray scattering and ensemble-optimization method |
SI | strong interactions |
SMFS | single molecule force spectroscopy |
WI | weak interactions |
WLC | worm-like-chain |
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Corti, R.; Marrano, C.A.; Salerno, D.; Brocca, S.; Natalello, A.; Santambrogio, C.; Legname, G.; Mantegazza, F.; Grandori, R.; Cassina, V. Depicting Conformational Ensembles of α-Synuclein by Single Molecule Force Spectroscopy and Native Mass Spectroscopy. Int. J. Mol. Sci. 2019, 20, 5181. https://doi.org/10.3390/ijms20205181
Corti R, Marrano CA, Salerno D, Brocca S, Natalello A, Santambrogio C, Legname G, Mantegazza F, Grandori R, Cassina V. Depicting Conformational Ensembles of α-Synuclein by Single Molecule Force Spectroscopy and Native Mass Spectroscopy. International Journal of Molecular Sciences. 2019; 20(20):5181. https://doi.org/10.3390/ijms20205181
Chicago/Turabian StyleCorti, Roberta, Claudia A. Marrano, Domenico Salerno, Stefania Brocca, Antonino Natalello, Carlo Santambrogio, Giuseppe Legname, Francesco Mantegazza, Rita Grandori, and Valeria Cassina. 2019. "Depicting Conformational Ensembles of α-Synuclein by Single Molecule Force Spectroscopy and Native Mass Spectroscopy" International Journal of Molecular Sciences 20, no. 20: 5181. https://doi.org/10.3390/ijms20205181