title: Using leucine zipper to facilitate alpha-synuclein assembly.
abstract: The accumulation of filamentous alpha-synuclein (alpha-S) is associated with Parkinson's disease. It remains controversial as to the mode (antiparallel or parallel) of alpha-S self-assembly and whether an exact alignment of the central hydrophobic region is essential. In the present study, we performed in vitro assembly using alpha-S with or without the attachment of artificial leucine zippers (Zips) capable of forming either parallel or antiparallel coiled coils and included a spacer in one derivative. Results showed that Zips accelerate filament assembly in both the parallel and antiparallel fashions, that a precise alignment of the central hydrophobic region is not essential, and that the antiparallel pairs displayed the highest thioflavin T signals. More importantly, cells expressing Zip-fused alpha-S, but not alpha-S alone, formed alpha-S immunopositive and thioflavin S-positive inclusions in 7 days. The results suggest that alpha-S can assemble in both parallel and antiparallel modes but have a higher tendency to assemble in the latter mode and that cells overexpressing Zip-fused alpha-S may be used to screen alpha-S assembly inhibitors due to enhanced ability to form inclusions.
authors.affiliation: Department of Neuroscience, Mayo Clinic College of Medicine, 4500 San Pablo Rd., Jacksonville, FL 32224, USA.
authors.forename: Peizhou
authors.initials: P
authors.lastname: Jiang
authors.forename: Li-wen
authors.initials: LW
authors.lastname: Ko
authors.forename: Karen R
authors.initials: KR
authors.lastname: Jansen
authors.forename: Todd E
authors.initials: TE
authors.lastname: Golde
authors.forename: Shu-Hui
authors.initials: SH
authors.lastname: Yen
mesh_terms: D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics; D006801:Humans; D016350:Leucine Zippers / Q000502:physiology*; D017433:Protein Structure, Secondary / Q000187:drug effects*; D011994:Recombinant Proteins / Q000737:chemistry; D014161:Transduction, Genetic; D051844:alpha-Synuclein / Q000096:biosynthesis*
chemical_list: D011994:Recombinant Proteins; D051844:alpha-Synuclein