Using leucine zipper to facilitate α-synuclein assembly Peizhou Jiang * ,† , Li-wen Ko * , Karen R. Jansen * , Todd E. Golde * and Shu-Hui Yen * ,1 * Department of Neuroscience, Mayo Clinic College of Medicine, Jacksonville, Florida, USA; and † Department of Histology and Embryology, School of Basic Medical Science, Southern Medical University, Guangzhou, Guangdong, China 1 Correspondence: Department of Neuroscience, Mayo Clinic College of Medicine, 4500 San Pablo Rd., Jacksonville, FL, 32224, USA. E-mail: yen.shu-hui@mayo.edu The accumulation of filamentous α-synuclein (α-S) is associated with Parkinson’s disease. It remains controversial as to the mode (antiparallel or parallel) of α-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 α-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 α-S, but not α-S alone, formed α-S immunopositive and thioflavin S-positive inclusions in 7 days. The results suggest that α-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 α-S may be used to screen α-S assembly inhibitors due to enhanced ability to form inclusions.—Jiang, P., Ko, L., Jansen, K. R., Golde, T. E., Yen, S.-H. Using leucine zipper to facilitate α-synuclein assembly. Key Words: aggregation • adeno-associated virus
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Using leucine zipper to facilitate α-synuclein assembly
Jiang, Peizhou; Ko, Li-wen; Jansen, Karen R.; Golde, Todd E.; Yen, Shu-Hui
Follow The FASEB Journal
, Volume 22 (9): 3165
Fed of American Socs for Experimental Biology – Sep 1, 2008
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