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Hydrogen–deuterium exchange in vivo to measure turnover of an ALS-associated mutant SOD1 protein in spinal cord of mice

Authors

  • George W. Farr,

    1. Howard Hughes Medical Institute, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
    2. Department of Genetics, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
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  • Zheng Ying,

    1. Howard Hughes Medical Institute, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
    2. Department of Genetics, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
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  • Wayne A. Fenton,

    1. Department of Genetics, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
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  • Arthur L. Horwich

    Corresponding author
    1. Howard Hughes Medical Institute, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
    2. Department of Genetics, Yale University School of Medicine, Boyer Center, New Haven, Connecticut 06510
    • Department of Genetics/HHMI, Yale School of Medicine, Boyer Center, 295 Congress Ave., New Haven, CT 06510
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Abstract

Mutations of cytosolic Cu/Zn superoxide dismutase 1 (SOD1) in humans and overexpression of mutant human SOD1 genes in transgenic mice are associated with the motor neuron degenerative condition known as amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease). Gain-of-function toxicity from the mutant protein expressed in motor neurons, associated with its misfolding and aggregation, leads to dysfunction and cell death, associated with paralyzing disease. Here, using hydrogen–deuterium exchange in intact mice in vivo, we have addressed whether an ALS-associated mutant protein, G85R SOD1–YFP, is subject to the same rate of turnover in spinal cord both early in the course of the disease and later. We find that the mutant protein turns over about 10-fold faster than a similarly expressed wild-type fusion and that there is no significant change in the rate of turnover as animals age and disease progresses.

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