AUTHOR=Guzzo  Adrien , Delarue  Patrice , Rojas Ana , Nicolaï  Adrien , Maisuradze  Gia G. , Senet Patrick 

TITLE=Wild-Type α-Synuclein and Variants Occur in Different Disordered Dimers and Pre-Fibrillar Conformations in Early Stage of Aggregation

JOURNAL=Frontiers in Molecular Biosciences

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/molecular-biosciences/articles/10.3389/fmolb.2022.910104

DOI=10.3389/fmolb.2022.910104

ISSN=2296-889X

ABSTRACT=<p><italic>α</italic>-Synuclein is a 140 amino-acid intrinsically disordered protein mainly found in the brain. Toxic <italic>α</italic>-synuclein aggregates are the molecular hallmarks of Parkinson’s disease. <italic>In vitro</italic> studies showed that <italic>α</italic>-synuclein aggregates in oligomeric structures of several 10th of monomers and into cylindrical structures (fibrils), comprising hundred to thousands of proteins, with polymorphic cross-<italic>β</italic>-sheet conformations. Oligomeric species, formed at the early stage of aggregation remain, however, poorly understood and are hypothezised to be the most toxic aggregates. Here, we studied the formation of wild-type (WT) and mutant (A30P, A53T, and E46K) dimers of <italic>α</italic>-synuclein using coarse-grained molecular dynamics. We identified two principal segments of the sequence with a higher propensity to aggregate in the early stage of dimerization: residues 36–55 and residues 66–95. The transient <italic>α</italic>-helices (residues 53–65 and 73–82) of <italic>α</italic>-synuclein monomers are destabilized by A53T and E46K mutations, which favors the formation of fibril native contacts in the N-terminal region, whereas the helix 53–65 prevents the propagation of fibril native contacts along the sequence for the WT in the early stages of dimerization. The present results indicate that dimers do not adopt the Greek key motif of the monomer fold in fibrils but form a majority of disordered aggregates and a minority (9–15%) of pre-fibrillar dimers both with intra-molecular and intermolecular <italic>β</italic>-sheets. The percentage of residues in parallel <italic>β</italic>-sheets is by increasing order monomer <inline-formula id="inf1"><mml:math id="m1" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mo>&lt;</mml:mo></mml:math></inline-formula> disordered dimers <inline-formula id="inf2"><mml:math id="m2" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mo>&lt;</mml:mo></mml:math></inline-formula> pre-fibrillar dimers. Native fibril contacts between the two monomers are present in the NAC domain for WT, A30P, and A53T and in the N-domain for A53T and E46K. Structural properties of pre-fibrillar dimers agree with rupture-force atomic force microscopy and single-molecule Förster resonance energy transfer available data. This suggests that the pre-fibrillar dimers might correspond to the smallest type B toxic oligomers. The probability density of the dimer gyration radius is multi-peaks with an average radius that is 10 Å larger than the one of the monomers for all proteins. The present results indicate that even the elementary <italic>α</italic>-synuclein aggregation step, the dimerization, is a complicated phenomenon that does not only involve the NAC region.</p>