{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Shuster SO"],"funding":["Intramural NIH HHS","National Heart, Lung, and Blood Institute"],"pagination":["3781-3789"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC8080960"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["125(15)"],"pubmed_abstract":["Aggregated TAR DNA-binding protein 43 (TDP-43) forms the cytoplasmic hallmarks associated with patients suffering from amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin. Under normal conditions, TDP-43 is a 414-amino acid protein; however, aggregates are enriched with N-terminal truncations which contain residues 267-414, known as the C-terminal domain of TDP-43 (TDP-43<sub>CTD</sub>). To gain residue-specific information on the aggregation process of TDP-43<sub>CTD</sub>, we created three single-Trp containing mutants (W385F/W412F, W334F/W412F, and W334F/W385F) by substituting two of the three native Trp residues with Phe, yielding fluorescent probes at W334, W385, and W412, respectively. Aggregation kinetics, secondary structure, and fibril morphology were compared to the wild-type protein using thioflavin-T fluorescence, Raman spectroscopy, and transmission electron microscopy, respectively. While only W334 is determined to be in the proteinase-K resistant core, all three sites are sensitive reporters of aggregation, revealing site-specific differences. Interestingly, W334 exhibited unusual multistep Trp kinetics, pinpointing a distinctive role for W334 and its nearby region during aggregation. This behavior is retained even upon seeding, suggesting the observed spectral change is related to fibril growth. This work provides new insights into the aggregation mechanism of TDP-43<sub>CTD</sub> and exemplifies the advantages of Trp as a site-specific environmentally sensitive fluorescent probe."],"journal":["The journal of physical chemistry. B"],"pubmed_title":["Tryptophan Probes of TDP-43 C-Terminal Domain Amyloid Formation."],"pmcid":["PMC8080960"],"funding_grant_id":["ZIA HL001055"],"pubmed_authors":["Lee JC","Shuster SO"],"additional_accession":[]},"is_claimable":false,"name":"Tryptophan Probes of TDP-43 C-Terminal Domain Amyloid Formation.","description":"Aggregated TAR DNA-binding protein 43 (TDP-43) forms the cytoplasmic hallmarks associated with patients suffering from amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitin. Under normal conditions, TDP-43 is a 414-amino acid protein; however, aggregates are enriched with N-terminal truncations which contain residues 267-414, known as the C-terminal domain of TDP-43 (TDP-43<sub>CTD</sub>). To gain residue-specific information on the aggregation process of TDP-43<sub>CTD</sub>, we created three single-Trp containing mutants (W385F/W412F, W334F/W412F, and W334F/W385F) by substituting two of the three native Trp residues with Phe, yielding fluorescent probes at W334, W385, and W412, respectively. Aggregation kinetics, secondary structure, and fibril morphology were compared to the wild-type protein using thioflavin-T fluorescence, Raman spectroscopy, and transmission electron microscopy, respectively. While only W334 is determined to be in the proteinase-K resistant core, all three sites are sensitive reporters of aggregation, revealing site-specific differences. Interestingly, W334 exhibited unusual multistep Trp kinetics, pinpointing a distinctive role for W334 and its nearby region during aggregation. This behavior is retained even upon seeding, suggesting the observed spectral change is related to fibril growth. This work provides new insights into the aggregation mechanism of TDP-43<sub>CTD</sub> and exemplifies the advantages of Trp as a site-specific environmentally sensitive fluorescent probe.","dates":{"release":"2021-01-01T00:00:00Z","publication":"2021 Apr","modification":"2025-04-04T10:15:43.156Z","creation":"2025-04-04T10:15:43.156Z"},"accession":"S-EPMC8080960","cross_references":{"pubmed":["33835818"],"doi":["10.1021/acs.jpcb.1c00767"]}}