title: The interconversion between a flexible β-sheet and a fibril β-arrangement constitutes the main conformational event during misfolding of PSD95-PDZ3 domain.
abstract: The temperature-induced misfolding pathway of PDZ3, the third PDZ domain of the PSD95 neuronal protein, is populated by a trimeric β-sheet-rich intermediate state that leads to a stepwise and reversible formation of supramacromolecular structures. Using FTIR, we have found that misfolding of this pathway is not due to different ensembles of a variety of precursors, but comes mainly from the interconversion of a flexible β-sheet of the domain to wormlike fibrils. The appearance of the wormlike fibril FTIR component is also accompanied by a slight decrease of the band that corresponds to loops in the native state, whereas the rest of the regular elements of secondary structure are fairly well maintained upon misfolding. Transmission electron microscope micrographs have confirmed the presence of wormlike fibrils upon heating at 60°C, where the trimeric intermediate is maximally populated. Toxicity assays in the human neuroblastoma cell line SH-SY5Y show that cytotoxicity increases as the aggregation pathway proceeds. NMR analysis of chemical shifts as a function of temperature has revealed, as one of the main conformational aspects of such an interconversion at the residue level, that the β-sheet arrangement around strand β3 promotes the change that drives misfolding of the PDZ3 domain.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Marin-Argany
authors.forename: Adela M
authors.initials: AM
authors.lastname: Candel
authors.forename: Javier
authors.initials: J
authors.lastname: Murciano-Calles
authors.forename: Jose C
authors.initials: JC
authors.lastname: Martinez
authors.forename: Sandra
authors.initials: S
authors.lastname: Villegas
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000633:toxicity; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054731:PDZ Domains; D017510:Protein Folding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary
chemical_list: D008565:Membrane Proteins