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Amyloid-like filaments and water-filled nanotubes formed by SOD1 mutant proteins linked to familial ALS

Nature Structural & Molecular Biology volume 10, pages 461–467 (2003)Cite this article

Abstract

Mutations in the SOD1 gene cause the autosomal dominant, neurodegenerative disorder familial amyotrophic lateral sclerosis (FALS). In spinal cord neurons of human FALS patients and in transgenic mice expressing these mutant proteins, aggregates containing FALS SOD1 are observed. Accumulation of SOD1 aggregates is believed to interfere with axonal transport, protein degradation and anti-apoptotic functions of the neuronal cellular machinery. Here we show that metal-deficient, pathogenic SOD1 mutant proteins crystallize in three different crystal forms, all of which reveal higher-order assemblies of aligned β-sheets. Amyloid-like filaments and water-filled nanotubes arise through extensive interactions between loop and β-barrel elements of neighboring mutant SOD1 molecules. In all cases, non-native conformational changes permit a gain of interaction between dimers that leads to higher-order arrays. Normal β-sheet–containing proteins avoid such self-association by preventing their edge strands from making intermolecular interactions. Loss of this protection through conformational rearrangement in the metal-deficient enzyme could be a toxic property common to mutants of SOD1 linked to FALS.

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Figure 1: Disorder and conformational changes in pathogenic SOD1 molecules leading to GOI interfaces and filamentous assembly.
Figure 2: GOI interfaces in pathogenic SOD1 give rise to cross-β fibrils in two different crystal systems.
Figure 3: Two orthogonal views of apo H46R filament 2, represented by three SOD1 dimers in the same orientation as in Fig. 2a.
Figure 4: Gain-of-interaction in Zn–H46R SOD1 giving rise to water-filled helical filaments.

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References

  1. Haverkamp, L.J., Appel, V. & Appel, S.H. Natural history of amyotrophic lateral sclerosis in a database population. Validation of a scoring system and a model for survival prediction. Brain 118, 707–719 (1995).

    Article  Google Scholar 

  2. Deng, H.X. et al. Amyotrophic lateral sclerosis and structural defects in Cu,Zn superoxide dismutase. Science 261, 1047–1051 (1993).

    Article  CAS  Google Scholar 

  3. Rosen, D.R. et al. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 362, 59–62 (1993).

    Article  CAS  Google Scholar 

  4. Fridovich, I. Superoxide dismutases. An adaptation to a paramagnetic gas. J. Biol. Chem. 264, 7761–7764 (1989).

    CAS  PubMed  Google Scholar 

  5. Bowling, A.C., Schulz, J.B., Brown, R.H. Jr. & Beal, M.F. Superoxide dismutase activity, oxidative damage, and mitochondrial energy metabolism in familial and sporadic amyotrophic lateral sclerosis. J. Neurochem. 61, 2322–2325 (1993).

    Article  CAS  Google Scholar 

  6. Reaume, A.G. et al. Motor neurons in Cu/Zn superoxide dismutase-deficient mice develop normally but exhibit enhanced cell death after axonal injury. Nat. Genet. 13, 43–47 (1996).

    Article  CAS  Google Scholar 

  7. Ripps, M.E., Huntley, G.W., Hof, P.R., Morrison, J.H. & Gordon, J.W. Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis. Proc. Natl. Acad. Sci. USA 92, 689–693 (1995).

    Article  CAS  Google Scholar 

  8. Bruijn, L.I. et al. ALS-linked SOD1 mutant G85R mediates damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions. Neuron 18, 327–338 (1997).

    Article  CAS  Google Scholar 

  9. Gurney, M.E. et al. Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264, 1772–1775 (1994).

    Article  CAS  Google Scholar 

  10. Wong, P.C. et al. An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria. Neuron 14, 1105–1116 (1995).

    Article  CAS  Google Scholar 

  11. Wiedau-Pazos, M. et al. Altered reactivity of superoxide dismutase in familial amyotrophic lateral sclerosis. Science 271, 515–518 (1996).

    Article  CAS  Google Scholar 

  12. Yim, M.B. et al. A gain-of-function of an amyotrophic lateral sclerosis-associated Cu,Zn-superoxide dismutase mutant: an enhancement of free radical formation due to a decrease in Km for hydrogen peroxide. Proc. Natl. Acad. Sci. USA 93, 5709–5714 (1996).

    Article  CAS  Google Scholar 

  13. Beckman, J.S., Chen, J., Crow, J.P. & Ye, Y.Z. Reactions of nitric oxide, superoxide and peroxynitrite with superoxide dismutase in neurodegeneration. Prog. Brain Res. 103, 371–380 (1994).

    Article  CAS  Google Scholar 

  14. Estevez, A.G. et al. Induction of nitric oxide-dependent apoptosis in motor neurons by zinc-deficient superoxide dismutase. Science 286, 2498–2500 (1999).

    Article  CAS  Google Scholar 

  15. Bruijn, L.I. et al. Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1. Science 281, 1851–1854 (1998).

    Article  CAS  Google Scholar 

  16. Wang, J. et al. Fibrillar inclusions and motor neuron degeneration in transgenic mice expressing superoxide dismutase 1 with a disrupted copper-binding site. Neurobiol. Dis. 10, 128–138 (2002).

    Article  CAS  Google Scholar 

  17. Bruening, W. et al. Up-regulation of protein chaperones preserves viability of cells expressing toxic Cu/Zn-superoxide dismutase mutants associated with amyotrophic lateral sclerosis. J. Neurochem. 72, 693–699 (1999).

    Article  CAS  Google Scholar 

  18. Okado-Matsumoto, A. & Fridovich, I. Amyotrophic lateral sclerosis: a proposed mechanism. Proc. Natl. Acad. Sci. USA 99, 9010–9014 (2002).

    Article  CAS  Google Scholar 

  19. Borchelt, D.R. et al. Axonal transport of mutant superoxide dismutase 1 and focal axonal abnormalities in the proximal axons of transgenic mice. Neurobiol. Dis. 5, 27–35 (1998).

    Article  CAS  Google Scholar 

  20. Williamson, T.L. & Cleveland, D.W. Slowing of axonal transport is a very early event in the toxicity of ALS-linked SOD1 mutants to motor neurons. Nat. Neurosci. 2, 50–56 (1999).

    Article  CAS  Google Scholar 

  21. Johnston, J.A., Dalton, M.J., Gurney, M.E. & Kopito, R.R. Formation of high molecular weight complexes of mutant Cu, Zn-superoxide dismutase in a mouse model for familial amyotrophic lateral sclerosis. Proc. Natl. Acad. Sci. USA 97, 12571–12576 (2000).

    Article  CAS  Google Scholar 

  22. Hayward, L.J. et al. Decreased metallation and activity in subsets of mutant superoxide dismutases associated with familial amyotrophic lateral sclerosis. J. Biol. Chem. 277, 15923–15931 (2002).

    Article  CAS  Google Scholar 

  23. Liu, H. et al. Copper2+ binding to the surface residue cysteine 111 of His46Arg human copper-zinc superoxide dismutase, a familial amyotrophic lateral sclerosis mutant. Biochemistry 39, 8125–8132 (2000).

    Article  CAS  Google Scholar 

  24. Rodriguez, J.A. et al. Familial ALS-associated mutations decrease the thermal stability of distinctly metallated species of human copper-zinc superoxide dismutase. J. Biol. Chem. 277, 15932–15937 (2002).

    Article  CAS  Google Scholar 

  25. Lyons, T.J. et al. Mutations in copper-zinc superoxide dismutase that cause amyotrophic lateral sclerosis alter the zinc binding site and the redox behavior of the protein. Proc. Natl. Acad. Sci. USA 93, 12240–12244 (1996).

    Article  CAS  Google Scholar 

  26. Crow, J.P., Sampson, J.B., Zhuang, Y., Thompson, J.A. & Beckman, J.S. Decreased zinc affinity of amyotrophic lateral sclerosis-associated superoxide dismutase mutants leads to enhanced catalysis of tyrosine nitration by peroxynitrite. J. Neurochem. 69, 1936–1944 (1997).

    Article  CAS  Google Scholar 

  27. Lyons, T.J. et al. The metal binding properties of the zinc site of yeast copper-zinc superoxide dismutase: implications for amyotrophic lateral sclerosis. J. Biol. Inorg. Chem. 5, 189–203 (2000).

    Article  CAS  Google Scholar 

  28. Subramaniam, J.R. et al. Mutant SOD1 causes motor neuron disease independent of copper chaperone-mediated copper loading. Nat. Neurosci. 5, 301–307 (2002).

    Article  CAS  Google Scholar 

  29. Serag, A.A., Altenbach, C., Gingery, M., Hubbell, W.L. & Yeates, T.O. Arrangement of subunits and ordering of β-strands in an amyloid sheet. Nat. Struct. Biol. 9, 734–739 (2002).

    Article  CAS  Google Scholar 

  30. Connors, L.H., Richardson, A.M., Theberge, R. & Costello, C.E. Tabulation of transthyretin (TTR) variants as of 1/1/2000. Amyloid 7, 54–69 (2000).

    Article  CAS  Google Scholar 

  31. Richardson, J.S. & Richardson, D.C. Natural β-sheet proteins use negative design to avoid edge-to-edge aggregation. Proc. Natl. Acad. Sci. USA 99, 2754–2759 (2002).

    Article  CAS  Google Scholar 

  32. Watanabe, M. et al. Histological evidence of protein aggregation in mutant SOD1 transgenic mice and in amyotrophic lateral sclerosis neural tissues. Neurobiol. Dis. 8, 933–941 (2001).

    Article  CAS  Google Scholar 

  33. Strange, R.W. et al. The structure of metal deficient wild type human Cu,Zn superoxide dismutase and its relevance to familial amyotrophic lateral sclerosis. J. Mol. Biol. 328, 877–891 (2003).

    Article  CAS  Google Scholar 

  34. Perutz, M.F., Finch, J.T., Berriman, J. & Lesk, A. Amyloid fibers are water-filled nanotubes. Proc. Natl. Acad. Sci. USA 99, 5591–5595 (2002).

    Article  CAS  Google Scholar 

  35. Lashuel, H.A., Hartley, D., Petre, B.M., Walz, T. & Lansbury, P.T. Jr. Neurodegenerative disease: amyloid pores from pathogenic mutations. Nature 418, 291 (2002).

    Article  CAS  Google Scholar 

  36. Sherman, M.Y. & Goldberg, A.L. Cellular defenses against unfolded proteins: a cell biologist thinks about neurodegenerative diseases. Neuron 29, 15–32 (2001).

    Article  CAS  Google Scholar 

  37. Shibata, N. et al. Intense superoxide dismutase-1 immunoreactivity in intracytoplasmic hyaline inclusions of familial amyotrophic lateral sclerosis with posterior column involvement. J. Neuropathol. Exp. Neurol. 55, 481–490 (1996).

    Article  CAS  Google Scholar 

  38. Wang, J., Xu, G. & Borchelt, D.R. High molecular weight complexes of mutant superoxide dismutase 1: age-dependent and tissue-specific accumulation. Neurobiol. Dis. 9, 139–148 (2002).

    Article  CAS  Google Scholar 

  39. Taylor, J.P., Hardy, J. & Fischbeck, K.H. Toxic proteins in neurodegenerative disease. Science 296, 1991–1995 (2002).

    Article  CAS  Google Scholar 

  40. Goto, J.J. et al. Loss of in vitro metal ion binding specificity in mutant copper-zinc superoxide dismutases associated with familial amyotrophic lateral sclerosis. J. Biol. Chem. 275, 1007–1014 (2000).

    Article  CAS  Google Scholar 

  41. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 306–326 (1997).

    Google Scholar 

  42. Hart, P.J. et al. Subunit asymmetry in the three-dimensional structure of a human CuZnSOD mutant found in familial amyotrophic lateral sclerosis. Protein Sci. 7, 545–555 (1998).

    Article  CAS  Google Scholar 

  43. Navaza, J. & Saludjian, P. AMoRe: an automated molecular replacement program package. Methods Enzymol. 276, 581–594 (1997).

    Article  CAS  Google Scholar 

  44. Kissinger, C.R., Gehlhaar, D.K. & Fogel, D.B. Rapid automated molecular replacement by evolutionary search. Acta Crystallogr. D 55, 484–491 (1999).

    Article  CAS  Google Scholar 

  45. Vagin, A.A. & Teplyakov, A. MOLREP: an automated program for molecular replacement. J. Appl. Crystallogr. 30, 1022–1025 (1997).

    Article  CAS  Google Scholar 

  46. Murshudov, G.N., Vagin, A.A., Lebedev, A., Wilson, K.S. & Dodson, E.J. Efficient anisotropic refinement of macromolecular structures using FFT. Acta Crystallogr. D 55, 247–255 (1999).

    Article  CAS  Google Scholar 

  47. Brunger, A.T. et al. Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998).

    Article  CAS  Google Scholar 

  48. Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991).

    Article  Google Scholar 

  49. Winn, M.D., Isupov, M.N. & Murshudov, G.N. Use of TLS parameters to model anisotropic displacements in macromolecular refinement. Acta Crystallogr. D 57, 122–133 (2001).

    Article  CAS  Google Scholar 

  50. Hooft, R.W., Vriend, G., Sander, C. & Abola, E.E. Errors in protein structures. Nature 381, 272 (1996).

    Article  CAS  Google Scholar 

  51. Laskowski, R.A., McArthur, M.W., Moss, D.S. & Thornton, J.M. PROCHECK: a program to check the stereochemical quality of protein structures. J. Appl. Crystallogr. 26, 283–291 (1993).

    Article  CAS  Google Scholar 

  52. Cohen, G.E. ALIGN: a program to superimpose protein coordinates, accounting for insertions and deletions. J. Appl. Crystallogr. 30, 1160–1161 (1997).

    Article  CAS  Google Scholar 

  53. Kraulis, P.J. MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr. 24, 946–950 (1991).

    Article  Google Scholar 

  54. Esnouf, R.M. Further additions to MolScript version 1.4, including reading and contouring of electron-density maps. Acta Crystallogr. D 55, 938–940 (1999).

    Article  CAS  Google Scholar 

  55. Read, R.J. Improved Fourier coefficients for maps using phases from partial structures with errors. Acta Crystallogr. A 42, 140–149 (1986).

    Article  Google Scholar 

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Acknowledgements

We thank L. Flaks, and J. Berendzen for support at beamline X8-C at the NSLS, Brookhaven National Laboratory; D. Cascio and M. Hough for their interest and valuable discussions; S. Holloway for assistance with the illustrations and our colleagues who have offered comments during the preparation of this manuscript. This work was supported by the National Institutes of Health (L.J.H., J.S.V. and P.J.H.), the Robert A. Welch Foundation (P.J.H.), the ALS Association (L.J.H., J.S.V and P.J.H.), the MND Association (S.S.H.) and a predoctoral fellowship from the Association for the Advancement of Aging Research (J.S.E.). Funding from CCLRC and resources at Daresbury are also gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry and the Center for Biomolecular Structure Analysis, The University of Texas, Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, 78229-3900, Texas, USA

    Jennifer Stine Elam, Alexander B Taylor & P John Hart

  2. Department of Synchrotron Radiation, Molecular Biophysics Group, CCLRC Daresbury Laboratory, Warrington, WA44AD, Cheshire, UK

    Richard Strange, Svetlana Antonyuk & S Samar Hasnain

  3. Department of Chemistry and Biochemistry, University of California, Los Angeles, 90095, California, USA

    Peter A Doucette, Jorge A Rodriguez, Joan Selverstone Valentine & Todd O Yeates

  4. Department of Neurology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, 01655, Massachusetts, USA

    Lawrence J Hayward

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Elam, J., Taylor, A., Strange, R. et al. Amyloid-like filaments and water-filled nanotubes formed by SOD1 mutant proteins linked to familial ALS. Nat Struct Mol Biol 10, 461–467 (2003). https://doi.org/10.1038/nsb935

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