title: The influence of pathological mutations and proline substitutions in TDP-43 glycine-rich peptides on its amyloid properties and cellular toxicity.
abstract: TAR DNA-binding protein (TDP-43) was identified as the major ubiquitinated component deposited in the inclusion bodies in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U) in 2006. Later on, numerous ALS-related mutations were found in either the glycine or glutamine/asparagine-rich region on the TDP-43 C-terminus, which hinted on the importance of mutations on the disease pathogenesis. However, how the structural conversion was influenced by the mutations and the biological significance of these peptides remains unclear. In this work, various peptides bearing pathogenic or de novo designed mutations were synthesized and displayed their ability to form twisted amyloid fibers, cause liposome leakage, and mediate cellular toxicity as confirmed by transmission electron microscopy (TEM), circular dichroism (CD), Thioflavin T (ThT) assay, Raman spectroscopy, calcein leakage assay, and cell viability assay. We have also shown that replacing glycines with prolines, known to obstruct β-sheet formation, at the different positions in these peptides may influence the amyloidogenesis process and neurotoxicity. In these cases, GGG308PPP mutant was not able to form beta-amyloid, cause liposome leakage, nor jeopardized cell survival, which hinted on the importance of the glycines (308-310) during amyloidogenesis.
authors.affiliation: Taiwan International Graduate Program in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan; Institute of Chemistry, Academia Sinica, Taipei, Taiwan; Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan; Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Chia-Sui
authors.initials: CS
authors.lastname: Sun
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei, Taiwan; Department of Chemistry, National Taiwan University, Taipei, Taiwan.
authors.forename: Cindy Yu-Hsiang
authors.initials: CY
authors.lastname: Wang
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
authors.forename: Bryan Po-Wen
authors.initials: BP
authors.lastname: Chen
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
authors.forename: Ruei-Yu
authors.initials: RY
authors.lastname: He
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei, Taiwan; Department of Chemistry, National Taiwan University, Taipei, Taiwan.
authors.forename: Gerard Chun-Hao
authors.initials: GC
authors.lastname: Liu
authors.affiliation: Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan.
authors.forename: Chih-Hsien
authors.initials: CH
authors.lastname: Wang
authors.affiliation: Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan.
authors.forename: Wenlung
authors.initials: W
authors.lastname: Chen
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
authors.forename: Yijuang
authors.initials: Y
authors.lastname: Chern
authors.affiliation: Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
authors.forename: Joseph Jen-Tse
authors.initials: JJ
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000682:Amyloid / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D052160:Benzothiazoles; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000633:toxicity; D005998:Glycine / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism / Q000633:toxicity; D009154:Mutation / Q000235:genetics*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D011392:Proline / Q000235:genetics*; D066329:Protein Aggregates / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D017433:Protein Structure, Secondary; D013059:Spectrum Analysis, Raman; D013844:Thiazoles / Q000378:metabolism; D013997:Time Factors
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D004268:DNA-Binding Proteins; D050505:Mutant Proteins; D010455:Peptides; D066329:Protein Aggregates; D013844:Thiazoles; C009462:thioflavin T; D011392:Proline; D005998:Glycine
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