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|Title:||Conformational dynamics of alpha-synuclein: insights from mass spectrometry|
|Author:||Phillips, Ashley S.|
Gomes, Alexandre F.
Kalapothakis, Jason M. D.
Gillam, Jay E.
Gozzo, Fabio C.
Barran, Perdita E.
|Abstract:||The aggregation and deposition of alpha-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 alpha-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 similar to 1600 angstrom(2)). 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 (similar to 1200 angstrom(2)), Extended (similar to 1500 angstrom(2)) and Unfolded (similar to 2350 angstrom(2)) 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 alpha-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 alpha-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|
|Subject:||Doença de Parkinson|
|Editor:||Royal Society of Chemistry|
|Appears in Collections:||IQ - Artigos e Outros Documentos|
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