TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease

Barbara Baldo; Paolo Paganetti; Stephan Grueninger; David Marcellin; Linda S. Kaltenbach; Donald C. Lo; Martin Semmelroth; Andjelija Zivanovic; Dorothee Abramowski; Donna Smith; Gregor P. Lotz; Gillian P. Bates; Andreas Weiss

doi:10.1016/j.chembiol.2011.12.020

TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease

Barbara Baldo, Paolo Paganetti, Stephan Grueninger, David Marcellin, Linda S. Kaltenbach, Donald C. Lo, Martin Semmelroth, Andjelija Zivanovic, Dorothee Abramowski, Donna Smith, Gregor P. Lotz, Gillian P. Bates, Andreas Weiss

Medical & Molecular Genetics

Research output: Contribution to journal › Article › peer-review

64 Citations (Scopus)

Abstract

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.

Original language	English
Pages (from-to)	264 - 275
Number of pages	12
Journal	Chemistry and Biology
Volume	19
Issue number	2
DOIs	https://doi.org/10.1016/j.chembiol.2011.12.020
Publication status	Published - 24 Feb 2012

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10.1016/j.chembiol.2011.12.020

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Baldo, B., Paganetti, P., Grueninger, S., Marcellin, D., Kaltenbach, L. S., Lo, D. C., Semmelroth, M., Zivanovic, A., Abramowski, D., Smith, D., Lotz, G. P., Bates, G. P., & Weiss, A. (2012). TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease. Chemistry and Biology, 19(2), 264 - 275. https://doi.org/10.1016/j.chembiol.2011.12.020

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title = "TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease",

abstract = "Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.",

author = "Barbara Baldo and Paolo Paganetti and Stephan Grueninger and David Marcellin and Kaltenbach, {Linda S.} and Lo, {Donald C.} and Martin Semmelroth and Andjelija Zivanovic and Dorothee Abramowski and Donna Smith and Lotz, {Gregor P.} and Bates, {Gillian P.} and Andreas Weiss",

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doi = "10.1016/j.chembiol.2011.12.020",

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Baldo, B, Paganetti, P, Grueninger, S, Marcellin, D, Kaltenbach, LS, Lo, DC, Semmelroth, M, Zivanovic, A, Abramowski, D, Smith, D, Lotz, GP, Bates, GP & Weiss, A 2012, 'TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease', Chemistry and Biology, vol. 19, no. 2, pp. 264 - 275. https://doi.org/10.1016/j.chembiol.2011.12.020

TY - JOUR

T1 - TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease

AU - Baldo, Barbara

AU - Paganetti, Paolo

AU - Grueninger, Stephan

AU - Marcellin, David

AU - Kaltenbach, Linda S.

AU - Lo, Donald C.

AU - Semmelroth, Martin

AU - Zivanovic, Andjelija

AU - Abramowski, Dorothee

AU - Smith, Donna

AU - Lotz, Gregor P.

AU - Bates, Gillian P.

AU - Weiss, Andreas

PY - 2012/2/24

Y1 - 2012/2/24

N2 - Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.

AB - Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.

U2 - 10.1016/j.chembiol.2011.12.020

DO - 10.1016/j.chembiol.2011.12.020

M3 - Article

VL - 19

SP - 264

EP - 275

JO - Chemistry and Biology

JF - Chemistry and Biology

IS - 2

ER -

TR-FRET-Based Duplex Immunoassay Reveals an Inverse Correlation of Soluble and Aggregated Mutant huntingtin in Huntington's Disease

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