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bioRxiv · 10.1101/2024.05.01.592011

Amyloid Accelerator Polyphosphate Implicated as the Mystery Density in α-Synuclein Fibrils

Huettemann, P.·Mahadevan, P.·Lempart, J.·Tse, E.·Dehury, B.·Edwards, B. F.·Southworth, D. R.·Sahoo, B. R.·Jakob, U.
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Abstract

Aberrant aggregation of -Synuclein is the pathological hallmark of a set of neurodegenerative diseases termed synucleinopathies. Recent advances in cryo-electron microscopy have led to the structural determination of the first synucleinopathy-derived -Synuclein fibrils, which contain a non-proteinaceous, "mystery density" at the core of the protofilaments, hypothesized to be highly negatively charged. Guided by previous studies that demonstrated that polyphosphate (polyP), a universally conserved polyanion, significantly accelerates -Synuclein fibril formation, we conducted blind docking and molecular dynamics simulation experiments to model the polyP binding site in -Synuclein fibrils. Here we demonstrate that our models uniformly place polyP into the lysine-rich pocket, which coordinates the mystery density in patient-derived fibrils. Subsequent in vitro studies and experiments in cells revealed that substitution of the two critical lysine residues K43 and K45 leads to a loss of all previously reported effects of polyP binding on -Synuclein, including stimulation of fibril formation, change in filament conformation and stability as well as alleviation of cytotoxicity. In summary, our study demonstrates that polyP fits the unknown electron density present in in vivo -Synuclein fibrils and suggests that polyP exerts its functions by neutralizing charge repulsion between neighboring lysine residues.

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Huettemann, P., Mahadevan, P., Lempart, J., Tse, E., Dehury, B., Edwards, B. F., Southworth, D. R., Sahoo, B. R., Jakob, U.. 2024-05-02. Amyloid Accelerator Polyphosphate Implicated as the Mystery Density in α-Synuclein Fibrils. https://doi.org/10.1101/2024.05.01.592011

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