Detecting morphologically distinct oligomeric forms of alpha-synuclein.
Neuropathologic and genetics studies as well as transgenic animal models have provided strong evidence linking misfolding and aggregation of alpha-synuclein to the progression of Parkinson disease (PD) and other related disorders. A growing body of evidence implicates various oligomeric forms of alpha-synuclein as the toxic species responsible for neurodegeneration and neuronal cell death. Although numerous different oligomeric forms of alpha-synuclein have been identified in vitro, it is not known which forms are involved in PD or how, when, and where different forms contribute to the progression of PD. Reagents that can interact with specific aggregate forms of alpha-synuclein would be very useful not only as tools to study how different aggregate forms affect cell function, but also as potential diagnostic and therapeutic agents for PD. Here we show that a single chain antibody fragment (syn-10H scFv) isolated from a phage display antibody library binds to a larger, later stage oligomeric form of alpha-synuclein than a previously reported oligomeric specific scFv isolated in our laboratory. The scFv described here inhibits aggregation of alpha-synuclein in vitro, blocks extracellular alpha-synuclein-induced toxicity in both undifferentiated and differentiated human neuroblastoma cell lines (SH-SY5Y), and specifically recognizes naturally occurring aggregates in PD but not in healthy human brain tissue.
Emadi
Sharareh
S
Department of Chemical Engineering, Arizona State University, Tempe, Arizona 85287-6006, USA.
Kasturirangan
Srinath
S
Wang
Min S
MS
Schulz
Philip
P
Sierks
Michael R
MR
D001921:Brain / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002470:Cell Survival; D018450:Disease Progression; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry; D018625:Microscopy, Atomic Force; D010300:Parkinson Disease / Q000378:metabolism* / Q000473:pathology; D019151:Peptide Library; D011485:Protein Binding; D013997:Time Factors; D051844:alpha-Synuclein / Q000737:chemistry* / Q000378:metabolism
D007128:Immunoglobulin Fragments; D019151:Peptide Library; D051844:alpha-Synuclein