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A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease.

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A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease. / Nagarajan, Archana; Bodhicharla, Rakesh; Winter, Jody; Anbalagan, Charumathi; Morgan, Kevin; Searle, Mark; Nazir, Aamir; Adenle, Ademola; Fineberg, April; Brady, Declan; Vere, Kelly; Richens, Jo; O'Shea, Paul; Bell, David; de-Pomerai, David.

In: CNS & neurological disorders drug targets, 21.08.2015.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Nagarajan, A, Bodhicharla, R, Winter, J, Anbalagan, C, Morgan, K, Searle, M, Nazir, A, Adenle, A, Fineberg, A, Brady, D, Vere, K, Richens, J, O'Shea, P, Bell, D & de-Pomerai, D 2015, 'A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease.', CNS & neurological disorders drug targets. <http://www.ncbi.nlm.nih.gov/pubmed/26295817>

APA

Nagarajan, A., Bodhicharla, R., Winter, J., Anbalagan, C., Morgan, K., Searle, M., Nazir, A., Adenle, A., Fineberg, A., Brady, D., Vere, K., Richens, J., O'Shea, P., Bell, D., & de-Pomerai, D. (2015). A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease. CNS & neurological disorders drug targets. http://www.ncbi.nlm.nih.gov/pubmed/26295817

Vancouver

Nagarajan A, Bodhicharla R, Winter J, Anbalagan C, Morgan K, Searle M et al. A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease. CNS & neurological disorders drug targets. 2015 Aug 21.

Author

Nagarajan, Archana ; Bodhicharla, Rakesh ; Winter, Jody ; Anbalagan, Charumathi ; Morgan, Kevin ; Searle, Mark ; Nazir, Aamir ; Adenle, Ademola ; Fineberg, April ; Brady, Declan ; Vere, Kelly ; Richens, Jo ; O'Shea, Paul ; Bell, David ; de-Pomerai, David. / A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease. In: CNS & neurological disorders drug targets. 2015.

Bibtex

@article{22112db40bec45468cb022b99123aa64,
title = "A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease.",
abstract = "The aggregation of α-synuclein (Syn or S) to form insoluble fibrils is important in the pathogenesis of Parkinson's disease, but key risk factors remain ill-defined. We have developed Fluorescence Resonance Energy Transfer (FRET)-based assays for α-synuclein aggregation, using Green Fluorescent Protein variants Cerulean (C) or Venus (V), fused to each other (CV, VC) or to human synuclein (SC, SV etc). Bacterially expressed proteins were purified to homogeneity, and C-terminal fusions SC and SV largely retained their ability to aggregate in vitro. FRET signals from mixtures of SC and SV were used to monitor aggregation. These fusion genes were linked to the C. elegans unc-54 myosin promoter to generate integrated transgenic strains. Increased FRET signals, indicative of S aggregation, were observed following treatment of unc-54::SC + unc-54::SV double transgenic worms with low concentrations of mercury or chlorpyrifos, or with RNAi against hsp-70 and hip-1. Opposite changes in Yellow Fluorescent Protein (YFP) fluorescence in an unc-54::SV strain (NL5901) are likely to reflect FRET from Yellow Fluorescent Protein to aggregates of Syn fusion protein. This could provide the basis for a high throughput screening assay, which could be used for studying the effects of toxic chemicals and environmental pollutants on the aggregation of proteins such as Syn in vivo.",
author = "Archana Nagarajan and Rakesh Bodhicharla and Jody Winter and Charumathi Anbalagan and Kevin Morgan and Mark Searle and Aamir Nazir and Ademola Adenle and April Fineberg and Declan Brady and Kelly Vere and Jo Richens and Paul O'Shea and David Bell and David de-Pomerai",
year = "2015",
month = aug,
day = "21",
language = "English",
journal = "CNS & neurological disorders drug targets",
issn = "1996-3181",
publisher = "Bentham Science Publishers B.V.",

}

RIS

TY - JOUR

T1 - A Fluorescence Resonance Energy Transfer Assay For Monitoring α-Synclein Aggregation In A Caenorhabditis elegans Model For Parkinson's Disease.

AU - Nagarajan, Archana

AU - Bodhicharla, Rakesh

AU - Winter, Jody

AU - Anbalagan, Charumathi

AU - Morgan, Kevin

AU - Searle, Mark

AU - Nazir, Aamir

AU - Adenle, Ademola

AU - Fineberg, April

AU - Brady, Declan

AU - Vere, Kelly

AU - Richens, Jo

AU - O'Shea, Paul

AU - Bell, David

AU - de-Pomerai, David

PY - 2015/8/21

Y1 - 2015/8/21

N2 - The aggregation of α-synuclein (Syn or S) to form insoluble fibrils is important in the pathogenesis of Parkinson's disease, but key risk factors remain ill-defined. We have developed Fluorescence Resonance Energy Transfer (FRET)-based assays for α-synuclein aggregation, using Green Fluorescent Protein variants Cerulean (C) or Venus (V), fused to each other (CV, VC) or to human synuclein (SC, SV etc). Bacterially expressed proteins were purified to homogeneity, and C-terminal fusions SC and SV largely retained their ability to aggregate in vitro. FRET signals from mixtures of SC and SV were used to monitor aggregation. These fusion genes were linked to the C. elegans unc-54 myosin promoter to generate integrated transgenic strains. Increased FRET signals, indicative of S aggregation, were observed following treatment of unc-54::SC + unc-54::SV double transgenic worms with low concentrations of mercury or chlorpyrifos, or with RNAi against hsp-70 and hip-1. Opposite changes in Yellow Fluorescent Protein (YFP) fluorescence in an unc-54::SV strain (NL5901) are likely to reflect FRET from Yellow Fluorescent Protein to aggregates of Syn fusion protein. This could provide the basis for a high throughput screening assay, which could be used for studying the effects of toxic chemicals and environmental pollutants on the aggregation of proteins such as Syn in vivo.

AB - The aggregation of α-synuclein (Syn or S) to form insoluble fibrils is important in the pathogenesis of Parkinson's disease, but key risk factors remain ill-defined. We have developed Fluorescence Resonance Energy Transfer (FRET)-based assays for α-synuclein aggregation, using Green Fluorescent Protein variants Cerulean (C) or Venus (V), fused to each other (CV, VC) or to human synuclein (SC, SV etc). Bacterially expressed proteins were purified to homogeneity, and C-terminal fusions SC and SV largely retained their ability to aggregate in vitro. FRET signals from mixtures of SC and SV were used to monitor aggregation. These fusion genes were linked to the C. elegans unc-54 myosin promoter to generate integrated transgenic strains. Increased FRET signals, indicative of S aggregation, were observed following treatment of unc-54::SC + unc-54::SV double transgenic worms with low concentrations of mercury or chlorpyrifos, or with RNAi against hsp-70 and hip-1. Opposite changes in Yellow Fluorescent Protein (YFP) fluorescence in an unc-54::SV strain (NL5901) are likely to reflect FRET from Yellow Fluorescent Protein to aggregates of Syn fusion protein. This could provide the basis for a high throughput screening assay, which could be used for studying the effects of toxic chemicals and environmental pollutants on the aggregation of proteins such as Syn in vivo.

M3 - Journal article

C2 - 26295817

JO - CNS & neurological disorders drug targets

JF - CNS & neurological disorders drug targets

SN - 1996-3181

ER -