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Rapid Cytotoxicity Screening Platform for Amyloid Inhibitors Using a Membrane-Potential Sensitive Fluorescent Probe
journal contribution
posted on 2013-01-02, 00:00 authored by Jihoon Kim, Yasuhiko Sasaki, Wataru Yoshida, Natsuki Kobayashi, Anthony J. Veloso, Kagan Kerman, Kazunori Ikebukuro, Koji SodeThe growing interest in membrane interactions of amyloidogenic
proteins indicates that lipid binding and the regulation of membrane
potential are critical to the onset and progression of neurodegenerative
diseases such as Parkinson’s (PD), Alzheimer’s (AD),
and prion diseases. Advancing the understanding of this field requires
the application of varied biophysical and biological techniques designed
to probe the characteristics and underlying mechanisms of membrane-peptide
interactions. Therefore, the development of a rapid cytotoxicity evaluation
system using a membrane potential-sensitive bis-oxonol fluorescent
dye, DiBAC4(3) is reported here. The exposure of C-terminal truncated
α-synuclein 119 (α-Syn119) and amyloid-β1–42 (Aβ1–42) to U2-OS cell cultures resulted
in an immediate, significant, and concentration-dependent increase
in fluorescence response of DiBAC4(3). This response was strongly
correlated with the cytotoxicity of α-Syn119 and Aβ1–42 as determined by conventional CC8 and ATP assays.
Furthermore, the capacity of well-defined polyphenolic antioxidants
(i.e., pyrroloquinoline quinone (PQQ), baicalein, (−)-epigallocatechin-3-gallate
(EGCG), and myricetin) to mitigate amyloid-induced cytotoxicity was
evaluated using the developed biosensing system. We envisage that
this work would accelerate the development of a rapid and cost-effective
high-throughput screening platform in drug discovery for AD and PD.
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Keywords
cytotoxicity evaluation systemlipid bindingPQQmembrane interactionsAmyloid InhibitorsFluorescent ProbeThePDCC 8Rapid Cytotoxicity Screening Platformprion diseasesbiosensing systemfluorescence responseEGCGneurodegenerative diseasesATP assaysdrug discoveryamyloidogenic proteinspolyphenolic antioxidantsDiBACSynAD
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