Abstract
Many mutations confer one or more toxic function(s) on copper/zinc superoxide dismutase 1 (SOD1) that impair motor neuron viability and cause familial amyotrophic lateral sclerosis (FALS). Using a conformation-specific antibody that detects misfolded SOD1 (C4F6), we found that oxidized wild-type SOD1 and mutant SOD1 share a conformational epitope that is not present in normal wild-type SOD1. In a subset of human sporadic ALS (SALS) cases, motor neurons in the lumbosacral spinal cord were markedly C4F6 immunoreactive, indicating that an aberrant wild-type SOD1 species was present. Recombinant, oxidized wild-type SOD1 and wild-type SOD1 immunopurified from SALS tissues inhibited kinesin-based fast axonal transport in a manner similar to that of FALS-linked mutant SOD1. Our findings suggest that wild-type SOD1 can be pathogenic in SALS and identify an SOD1-dependent pathogenic mechanism common to FALS and SALS.
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Acknowledgements
We are grateful to J. Landers and P. Sapp for DNA sequencing analysis of the SALS cases employed in this study, L. Hayward, A. Tiwari and R.-J. Chain for help with expression of recombinant wild-type SOD1, S. Berth, A. Leitman and M. Saparauskaite for help with axoplasm vesicle transport assays, A. Kaminska and L. Molla for help with biochemical experiments in squid axoplasm, M. Prudencio and D. Borchelt for cell lysates containing SOD1 G93 mutants, C. Vanderburg, E. Tamrazian, A. Bialik and the Diabetes and Endocrinology Research Center (University of Massachusetts Medical Center) for assistance with immunohistochemistry, K. Fitch and the Massachusetts Alzheimer Disease Research Center (P50AG005134) for assistance with human tissue samples, J. Zitzewitz for C6A/C111S-SOD1 protein, A. Weiss for assistance with mice, K. Green at the University of Massachusetts Medical Center Proteomics and Mass Spectrometry Facility for analysis of C6A/C111S-SOD1, and G. Petsko for insightful dialogue and support. We acknowledge financial support from the ALS Therapy Alliance-CVS Pharmacy (D.A.B. and G.M.), 2007/2008 Marine Biological Laboratory research fellowships (G.M.), the ALS Association (D.A.B., R.H.B. Jr, G.M. and S.T.B.), the US National Institutes of Health (D.A.B. (National Institute on Neurological Disorders and Stroke, RO1NS067206-01), R.H.B. Jr (National Institute on Neurological Disorders and Stroke, U01NS05225-03, R01NS050557-05, 1RC1NS068391-01 and 1RC2NS070342-01), S.T.B. and J.N.A.), Canadian Institutes of Health Research (J.-P.J.), the Angel Fund (R.H.B. Jr) and Project ALS (R.H.B. Jr).
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D.A.B., G.M., S.T.B. and R.H.B. Jr wrote the manuscript. D.A.B. prepared recombinant and immunopurified SOD1 proteins. G.M., Y.S. and S.T.B. performed vesicle motility assays and biochemical experiments in isolated squid axoplasm. N.M.K. and J.N.A. performed the mass spectrometry. F.G.-L. and J.-P.J. prepared the mutant-specific antibodies. P.P. made the SOD1 exon deleted constructs. H.G., D.M.-Y. and M.P.F. provided human tissues for staining. D.A.B., B.A.F. and N.L. performed western analyses. All of the authors reviewed and edited the manuscript.
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Bosco, D., Morfini, G., Karabacak, N. et al. Wild-type and mutant SOD1 share an aberrant conformation and a common pathogenic pathway in ALS. Nat Neurosci 13, 1396–1403 (2010). https://doi.org/10.1038/nn.2660
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DOI: https://doi.org/10.1038/nn.2660
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