α-Synuclein (α-syn) has been identified as the major component of Lewy bodies that characterize neurodegenerative synucleinopathies, including Parkinson's disease. Overexpression of α-syn, and prefibrillar α-syn oligomers, has been implicated in these pathologies; therefore, prevention of prefibril accumulation, and inhibition of other aberrant effects of overexpressed α-syn, could provide novel treatments. Here, we have selected a human single-chan Fv (scFv) antibody, D10, that binds human monomeric wild-type α-syn. We demonstrate, by retargeting assays and coimmunoprecipitation, that the D10 scFv is a specific and efficient intracellular antibody (intrabody). By transfecting the D10 scFv gene into an HEK 293 cell line that overexpresses wild-type α-syn, we show that the D10 intrabody stabilizes detergent-soluble monomeric α-syn and inhibits the formation of detergent-insoluble high-molecular-weight α-syn species. In addition, the D10 intrabody ameliorates the decreased cell adhesion that characterizes the α-syn-overexpressing cells. Given the important role of α-syn pathology, and the facility with which intrabodies can be further engineered in vitro, anti-α-syn intrabodies may represent novel molecular therapeutics for synucleinopathies, with implications for other neurodegenerative disorders caused by misfolded accumulated proteins.
- Lewy body
- Parkinson's disease
ASJC Scopus subject areas
- Molecular Biology
A human single-chain Fv intrabody blocks aberrant cellular effects of overexpressed α-synuclein. / Zhou, Chun; Emadi, Sharareh; Sierks, Michael; Messer, Anne.In: Molecular Therapy, Vol. 10, No. 6, 12.2004, p. 1023-1031.
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