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Amyotrophic lateral sclerosis is a non-amyloid disease in which extensive misfolding of SOD1 is unique to the familial form

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Abstract

Amyotrophic lateral sclerosis (ALS) is a conformational disease in which misfolding and aggregation of proteins such as SOD1 (familial ALS) and TDP-43 (sporadic ALS) are central features. The conformations adopted by such proteins within motor neurons in affected patients are not well known. We have developed a novel conformation-specific antibody (USOD) targeted against SOD1 residues 42–48 that specifically recognizes SOD1 in which the beta barrel is unfolded. Use of this antibody, in conjunction with the previously described SEDI antibody that recognizes the SOD1 dimer interface, allows a detailed investigation of the in vivo conformation of SOD1 at the residue-specific level. USOD and SEDI immunohistochemistry of spinal cord sections from ALS cases resulting from SOD1 mutations (A4V and ΔG27/P28) shows that inclusions within remaining motor neurons contain SOD1 with both an unfolded beta barrel and a disrupted dimer interface. Misfolded SOD1 can also be immunoprecipitated from spinal cord extracts of these cases using USOD. However, in ten cases of sporadic ALS, misfolded SOD1 is not detected by either immunohistochemistry or immunoprecipitation. Using the amyloid-specific dyes, Congo Red and Thioflavin S, we find that SOD1-positive inclusions in familial ALS, as well as TDP-43- and ubiquitin-positive inclusions in sporadic ALS, contain non-amyloid protein deposits. We conclude that SOD1 misfolding is not a feature of sporadic ALS, and that both SOD1-ALS and sporadic ALS, rather than being amyloid diseases, are conformational diseases that involve amorphous aggregation of misfolded protein. This knowledge will provide new insights into subcellular events that cause misfolding, aggregation and toxicity.

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Abbreviations

AD:

Alzheimer’s disease

ALS:

Amyotrophic lateral sclerosis

sALS:

Sporadic amyotrophic lateral sclerosis

CR:

Congo Red

CBD:

Corticobasal degeneration

DTT:

Dithiothreitol

ELISA:

Enzyme-linked immunosorbent assay

GuHCl:

Guanidine hydrochloride

MALDI:

Matrix-assisted laser desorption/ionization

PBS:

Phosphate-buffered saline

SOD1:

Superoxide dismutase 1

TDP-43:

TAR DNA binding protein-43

ThS:

Thioflavin S

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Acknowledgments

This study was supported by grants from the Canadian Institutes of Health Research to A.C. and J.R. We thank Nancy F.L. Ng for critical comments, and Kelvin So and Milan Ganguly for performing immunohistochemistry experiments.

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Authors and Affiliations

  1. Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, TMDT 4-305, 101 College Street, Toronto, ON, M5G 1L7, Canada

    Aaron Kerman & Avijit Chakrabartty

  2. Department of and Biochemistry, Ontario Cancer Institute, University of Toronto, TMDT 4-305, 101 College Street, Toronto, ON, M5G 1L7, Canada

    Aaron Kerman & Avijit Chakrabartty

  3. Department of Laboratory Medicine and Pathobiology, Centre for Research in Neurodegenerative Diseases, University of Toronto, 6 Queen’s Park Cres. W., Toronto, ON, M5S 3H2, Canada

    Hsueh-Ning Liu, Ekaterina Rogaeva & Janice Robertson

  4. Department of Laboratory Medicine and Pathobiology, UHN Path 11E426 Toronto General Hospital, University of Toronto, 200 Elizabeth St, Toronto, ON, M5G 2C4, Canada

    Sidney Croul

  5. Sunnybrook Health Sciences Centre, Toronto, ON, Canada

    Juan Bilbao & Lorne Zinman

Authors
  1. Aaron Kerman
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  2. Hsueh-Ning Liu
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  8. Avijit Chakrabartty
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Corresponding authors

Correspondence to Janice Robertson or Avijit Chakrabartty.

Additional information

A. Kerman and H.-N. Liu contributed equally to this work.

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Kerman, A., Liu, HN., Croul, S. et al. Amyotrophic lateral sclerosis is a non-amyloid disease in which extensive misfolding of SOD1 is unique to the familial form. Acta Neuropathol 119, 335–344 (2010). https://doi.org/10.1007/s00401-010-0646-5

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  • DOI: https://doi.org/10.1007/s00401-010-0646-5

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