Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a
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 amy...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-12-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/23/15291 |
| _version_ | 1797463038968725504 |
|---|---|
| author | Tanguy Leyder Julien Mignon Denis Mottet Catherine Michaux |
| author_facet | Tanguy Leyder Julien Mignon Denis Mottet Catherine Michaux |
| author_sort | Tanguy Leyder |
| collection | DOAJ |
| description | 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<sup>2+</sup>, Mg<sup>2+</sup>, Ni<sup>2+</sup>, and Zn<sup>2+</sup>, 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<sup>2+</sup> and Ni<sup>2+</sup> have a strong inhibitory effect on the aggregation of both isoforms, whereas Mg<sup>2+</sup> impedes C-TERb fibrillation and, on the contrary, enhances that of C-TERa. Upon Zn<sup>2+</sup> 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<sup>2+</sup> and Zn<sup>2+</sup> cations. |
| first_indexed | 2024-03-09T17:45:01Z |
| format | Article |
| id | doaj.art-e9e60a3fd54d427dafd323ed282bdb90 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T17:45:01Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e9e60a3fd54d427dafd323ed282bdb902023-11-24T11:16:32ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123231529110.3390/ijms232315291Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3aTanguy Leyder0Julien Mignon1Denis Mottet2Catherine Michaux3Laboratoire de Chimie Physique des Biomolécules, UCPTS, University of Namur, 61 rue de Bruxelles, 5000 Namur, BelgiumLaboratoire de Chimie Physique des Biomolécules, UCPTS, University of Namur, 61 rue de Bruxelles, 5000 Namur, BelgiumGene Expression and Cancer Laboratory, GIGA-Molecular Biology of Diseases, University of Liège, B34, Avenue de l’Hôpital, 4000 Liège, BelgiumLaboratoire de Chimie Physique des Biomolécules, UCPTS, University of Namur, 61 rue de Bruxelles, 5000 Namur, BelgiumDouble-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<sup>2+</sup>, Mg<sup>2+</sup>, Ni<sup>2+</sup>, and Zn<sup>2+</sup>, 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<sup>2+</sup> and Ni<sup>2+</sup> have a strong inhibitory effect on the aggregation of both isoforms, whereas Mg<sup>2+</sup> impedes C-TERb fibrillation and, on the contrary, enhances that of C-TERa. Upon Zn<sup>2+</sup> 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<sup>2+</sup> and Zn<sup>2+</sup> cations.https://www.mdpi.com/1422-0067/23/23/15291double-PHD fingers 3 (DPF3)intrinsically disordered proteinneurodegenerative diseasesaggregationamyloid fibrillationmetal cations |
| spellingShingle | Tanguy Leyder Julien Mignon Denis Mottet Catherine Michaux Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a International Journal of Molecular Sciences double-PHD fingers 3 (DPF3) intrinsically disordered protein neurodegenerative diseases aggregation amyloid fibrillation metal cations |
| title | Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a |
| title_full | Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a |
| title_fullStr | Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a |
| title_full_unstemmed | Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a |
| title_short | Unveiling the Metal-Dependent Aggregation Properties of the C-terminal Region of Amyloidogenic Intrinsically Disordered Protein Isoforms DPF3b and DPF3a |
| title_sort | unveiling the metal dependent aggregation properties of the c terminal region of amyloidogenic intrinsically disordered protein isoforms dpf3b and dpf3a |
| topic | double-PHD fingers 3 (DPF3) intrinsically disordered protein neurodegenerative diseases aggregation amyloid fibrillation metal cations |
| url | https://www.mdpi.com/1422-0067/23/23/15291 |
| work_keys_str_mv | AT tanguyleyder unveilingthemetaldependentaggregationpropertiesofthecterminalregionofamyloidogenicintrinsicallydisorderedproteinisoformsdpf3banddpf3a AT julienmignon unveilingthemetaldependentaggregationpropertiesofthecterminalregionofamyloidogenicintrinsicallydisorderedproteinisoformsdpf3banddpf3a AT denismottet unveilingthemetaldependentaggregationpropertiesofthecterminalregionofamyloidogenicintrinsicallydisorderedproteinisoformsdpf3banddpf3a AT catherinemichaux unveilingthemetaldependentaggregationpropertiesofthecterminalregionofamyloidogenicintrinsicallydisorderedproteinisoformsdpf3banddpf3a |