Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a

• Published in: International journal of molecular sciences 2022-12, Vol.23 (23), p.15291
• Main Authors: Leyder, Tanguy, Mignon, Julien, Mottet, Denis, Michaux, Catherine
• Format: Article
• Language: English
• Subjects: Aggregates; aggregation; Alzheimer's disease; Amino acids; Amyloid; Amyloid - metabolism; amyloid fibrillation; Amyloidogenesis; Binding; Biochemistry, biophysics & molecular biology; Biochimie, biophysique & biologie moléculaire; Cancer; Catalysis; Cations; Cations, Divalent; Chelating Agents; Chelating Agents - chemistry; Circular dichroism; Computer Science Applications; deep-blue autofluorescence; Dichroism; double-PHD fingers 3 (DPF3); Electron microscopy; Epigenetics; Fibrils; Filaments; General Medicine; Heart diseases; Homeostasis; Humans; Inorganic Chemistry; intrinsically disordered protein; Intrinsically Disordered Proteins; Intrinsically Disordered Proteins - chemistry; Isoforms; Life sciences; Magnesium; metal cations; Metal ions; Metal sheets; Metals; Molecular Biology; Morphology; Neurodegeneration; neurodegenerative diseases; Neurotoxicity; Organic Chemistry; Oxidative stress; Parkinson's disease; Physical and Theoretical Chemistry; Protein Isoforms; Protein structure; Proteins; Sciences du vivant; Spectroscopy; Tertiary structure; Zinc
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms232315291

Double-PHD fingers 3 (DPF3) is a BAF-associated human epigenetic regulator, which is increasingly recognised as a major contributor to various pathological contexts, such as cardiac defects, cancer, and neurodegenerative diseases. Recently, we unveiled that its two isoforms (DPF3b and DPF3a) are amyloidogenic intrinsically disordered proteins. DPF3 isoforms differ from their C-terminal region (C-TERb and C-TERa), containing zinc fingers and disordered domains. Herein, we investigated the disorder aggregation properties of C-TER isoforms. In agreement with the predictions, spectroscopy highlighted a lack of a highly ordered structure, especially for C-TERa. Over a few days, both C-TERs were shown to spontaneously assemble into similar antiparallel and parallel β-sheet-rich fibrils. Altered metal homeostasis being a neurodegeneration hallmark, we also assessed the influence of divalent metal cations, namely Cu , Mg , Ni , and Zn , on the C-TER aggregation pathway. Circular dichroism revealed that metal binding does not impair the formation of β-sheets, though metal-specific tertiary structure modifications were observed. Through intrinsic and extrinsic fluorescence, we found that metal cations differently affect C-TERb and C-TERa. Cu and Ni have a strong inhibitory effect on the aggregation of both isoforms, whereas Mg impedes C-TERb fibrillation and, on the contrary, enhances that of C-TERa. Upon Zn binding, C-TERb aggregation is also hindered, and the amyloid autofluorescence of C-TERa is remarkably red-shifted. Using electron microscopy, we confirmed that the metal-induced spectral changes are related to the morphological diversity of the aggregates. While metal-treated C-TERb formed breakable and fragmented filaments, C-TERa fibrils retained their flexibility and packing properties in the presence of Mg and Zn cations.