Abstract
Excessive aggregation of proteins has a major impact on cell fate and is a hallmark of amyloid diseases in humans. To resolve insoluble deposits and to maintain protein homeostasis, all cells use dedicated protein disaggregation, protein folding and protein degradation factors. Despite intense recent research, the underlying mechanisms controlling this key metabolic event are not well understood. Here, we analyzed how a single factor, the highly conserved serine protease HTRA1, degrades amyloid fibrils in an ATP-independent manner. This PDZ protease solubilizes protein fibrils and disintegrates the fibrillar core structure, allowing productive interaction of aggregated polypeptides with the active site for rapid degradation. The aggregate burden in a cellular model of cytoplasmic tau aggregation is thus reduced. Mechanistic aspects of ATP-independent proteolysis and its implications in amyloid diseases are discussed.
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Acknowledgements
We thank N. Schulze and M. Graessl for help with TIRFM, D. Grum and M. Merdanovic with ITC, S. Blaskowski with MS, V. Lux (University Duisburg-Essen) for providing purified PDZ domain and M. Breiden (University Duisburg-Essen) for providing tau. This work was supported by grants EH 100/14-1 (to M.E.) and SFB 1093 (to M.E., M.K. and B.S.) and INST 20876/127-1 FUGG (to M.K.) from Deutsche Forschungsgemeinschaft. The Research Institute of Molecular Pathology is funded by Boehringer Ingelheim.
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S.P. designed and carried out experiments, analyzed data and wrote the paper; A.S. and B.S. carried out AFM; F.K. and M.K. carried out MS; C.G. and S.R. carried out EM; T.C. performed biochemistry; and M.E. outlined the work and wrote the paper.
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Supplementary Results, Supplementary Table 1 and Supplementary Figures 1–9. (PDF 21400 kb)
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Uncut gel images: full gels of Figs 1a, 2c, 3a–c, 6a and 6e. (PDF 7454 kb)
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Poepsel, S., Sprengel, A., Sacca, B. et al. Determinants of amyloid fibril degradation by the PDZ protease HTRA1. Nat Chem Biol 11, 862–869 (2015). https://doi.org/10.1038/nchembio.1931
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DOI: https://doi.org/10.1038/nchembio.1931
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