Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association
Abstract TDP-43, a nucleocytoplasmic shuttle protein consisting of 414 residues, forms self-association in the nucleus for physiological gene regulation, while aggregation into amyloid (consisting of aberrant β-sheets) in the cytoplasm causes neurodegenerative diseases such as amyotrophic lateral sc...
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Nature Portfolio
2025-04-01
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| Series: | Communications Chemistry |
| Online Access: | https://doi.org/10.1038/s42004-025-01522-1 |
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| _version_ | 1850170882038693888 |
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| author | Daisuke Sasaki Mao Tenda Youhei Sohma |
| author_facet | Daisuke Sasaki Mao Tenda Youhei Sohma |
| author_sort | Daisuke Sasaki |
| collection | DOAJ |
| description | Abstract TDP-43, a nucleocytoplasmic shuttle protein consisting of 414 residues, forms self-association in the nucleus for physiological gene regulation, while aggregation into amyloid (consisting of aberrant β-sheets) in the cytoplasm causes neurodegenerative diseases such as amyotrophic lateral sclerosis. Post-translational phosphorylation of TDP-43 alters the self-association properties, which affects both the physiological function in the nucleus and the amyloidogenic potential in the cytoplasm, thereby impacting upon disease progression. However, insight into the role of per-residue phosphorylation in the self-association remains limited due to the difficulty in obtaining site-specifically phosphorylated TDP-43. Here, we demonstrate semi-synthesis of full-length TDP-43 that is uniformly phosphorylated at the 48th serine residue (designated as TDP1-414[pS48]). The synthetic scheme consisting of native chemical ligation followed by His-tag affinity chromatography efficiently gave TDP1-414(pS48) with a high purity. Interestingly, unlike non-phosphorylated TDP-43, the phosphorylated TDP-43 was found to have weak self-association property and to form aggregates that were not typical amyloid fibrils. Furthermore, chemical synthesis and three-dimensional structure analysis of the N-terminal domain (NTD, corresponding to TDP1-80) suggested that the phosphate ion at Ser48 weakens the inter-NTD interaction by inducing electrostatic repulsion. It significantly advances understanding of the pathological mechanisms involved in the post-translational modifications of TDP-43 associated with the neurodegenerative diseases. |
| format | Article |
| id | doaj-art-640a2ed1dafa4423829558c43f3a8532 |
| institution | OA Journals |
| issn | 2399-3669 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Chemistry |
| spelling | doaj-art-640a2ed1dafa4423829558c43f3a85322025-08-20T02:20:23ZengNature PortfolioCommunications Chemistry2399-36692025-04-018111010.1038/s42004-025-01522-1Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-associationDaisuke Sasaki0Mao Tenda1Youhei Sohma2Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Wakayama Medical UniversityDepartment of Medicinal Chemistry, School of Pharmaceutical Sciences, Wakayama Medical UniversityDepartment of Medicinal Chemistry, School of Pharmaceutical Sciences, Wakayama Medical UniversityAbstract TDP-43, a nucleocytoplasmic shuttle protein consisting of 414 residues, forms self-association in the nucleus for physiological gene regulation, while aggregation into amyloid (consisting of aberrant β-sheets) in the cytoplasm causes neurodegenerative diseases such as amyotrophic lateral sclerosis. Post-translational phosphorylation of TDP-43 alters the self-association properties, which affects both the physiological function in the nucleus and the amyloidogenic potential in the cytoplasm, thereby impacting upon disease progression. However, insight into the role of per-residue phosphorylation in the self-association remains limited due to the difficulty in obtaining site-specifically phosphorylated TDP-43. Here, we demonstrate semi-synthesis of full-length TDP-43 that is uniformly phosphorylated at the 48th serine residue (designated as TDP1-414[pS48]). The synthetic scheme consisting of native chemical ligation followed by His-tag affinity chromatography efficiently gave TDP1-414(pS48) with a high purity. Interestingly, unlike non-phosphorylated TDP-43, the phosphorylated TDP-43 was found to have weak self-association property and to form aggregates that were not typical amyloid fibrils. Furthermore, chemical synthesis and three-dimensional structure analysis of the N-terminal domain (NTD, corresponding to TDP1-80) suggested that the phosphate ion at Ser48 weakens the inter-NTD interaction by inducing electrostatic repulsion. It significantly advances understanding of the pathological mechanisms involved in the post-translational modifications of TDP-43 associated with the neurodegenerative diseases.https://doi.org/10.1038/s42004-025-01522-1 |
| spellingShingle | Daisuke Sasaki Mao Tenda Youhei Sohma Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association Communications Chemistry |
| title | Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association |
| title_full | Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association |
| title_fullStr | Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association |
| title_full_unstemmed | Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association |
| title_short | Semi-synthesis of TDP-43 reveals the effects of phosphorylation in N-terminal domain on self-association |
| title_sort | semi synthesis of tdp 43 reveals the effects of phosphorylation in n terminal domain on self association |
| url | https://doi.org/10.1038/s42004-025-01522-1 |
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