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Issue 9, 2015
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Conformational dynamics of α-synuclein: insights from mass spectrometry

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Abstract

The aggregation and deposition of α-synuclein in Lewy bodies is associated with the progression of Parkinson's disease. Here, Mass Spectrometry (MS) is used in combination with Ion Mobility (IM), chemical crosslinking and Electron Capture Dissociation (ECD) to probe transient structural elements of α-synuclein and its oligomers. Each of these reveals different aspects of the conformational heterogeneity of this 14 kDa protein. IM-MS analysis indicates that this protein is highly disordered, presenting in positive ionisation mode with a charge state range of 5 ≤ z ≤ 21 for the monomer, along with a collision cross section range of ∼1600 Å2. Chemical crosslinking applied in conjunction with IM-MS captures solution phase conformational families enabling comparison with those exhibited in the gas phase. Crosslinking IM-MS identifies 3 distinct conformational families, Compact (∼1200 Å2), Extended (∼1500 Å2) and Unfolded (∼2350 Å2) which correlate with those observed in solution. ECD-Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry (ECD-FT-ICR MS) highlights the effect of pH on α-synuclein structure, identifying the conformational flexibility of the N and C termini as well as providing evidence for structure in the core and at times the C terminus. A hypothesis is proposed for the variability displayed in the structural rearrangement of α-synuclein following changes in solution pH. Following a 120 h aggregation time course, we observe an increase in the ratio of dimer to monomer, but no gross conformational changes in either, beyond the significant variations that are observed day-to-day from this conformationally dynamic protein.

Graphical abstract: Conformational dynamics of α-synuclein: insights from mass spectrometry

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Publication details

The article was received on 16 Dec 2014, accepted on 25 Feb 2015 and first published on 25 Feb 2015


Article type: Paper
DOI: 10.1039/C4AN02306D
Citation: Analyst, 2015,140, 3070-3081
  • Open access: Creative Commons BY license
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    Conformational dynamics of α-synuclein: insights from mass spectrometry

    A. S. Phillips, A. F. Gomes, J. M. D. Kalapothakis, J. E. Gillam, J. Gasparavicius, F. C. Gozzo, T. Kunath, C. MacPhee and P. E. Barran, Analyst, 2015, 140, 3070
    DOI: 10.1039/C4AN02306D

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