Open AccessBifunctional Anti-Non-Amyloid Component α-Synuclein Nanobodies Are Protective In Situ
Author(s) -
David Butler,
Shubhada N. Joshi,
Erwin De Genst,
Ankit Baghel,
Christopher M. Dobson,
Anne Messer
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0165964
Subject(s) - synucleinopathies , alpha synuclein , huntingtin , chemistry , amyloid (mycology) , microbiology and biotechnology , mutant , bifunctional , biochemistry , biology , biophysics , parkinson's disease , inorganic chemistry , gene , catalysis , medicine , disease , pathology
Misfolding, abnormal accumulation, and secretion of α-Synuclein (α-Syn) are closely associated with synucleinopathies, including Parkinson’s disease (PD). VH14 is a human single domain intrabody selected against the non-amyloid component (NAC) hydrophobic interaction region of α-Syn, which is critical for initial aggregation. Using neuronal cell lines, we show that as a bifunctional nanobody fused to a proteasome targeting signal, VH14PEST can counteract heterologous proteostatic effects of mutant α-Syn on mutant huntingtin Exon1 and protect against α-Syn toxicity using propidium iodide or Annexin V readouts. We compared this anti-NAC candidate to NbSyn87, which binds to the C-terminus of α-Syn. NbSyn87PEST degrades α-Syn as well or better than VH14PEST. However, while both candidates reduced toxicity, VH14PEST appears more effective in both proteostatic stress and toxicity assays. These results show that the approach of reducing intracellular monomeric targets with novel antibody engineering technology should allow in vivo modulation of proteostatic pathologies.