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

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.

[1]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[2]  C. Costello,et al.  Sequence Communication: Tabulation of transthyretin (TTR) variants as of 1/1/2000 , 2000, Amyloid : the international journal of experimental and clinical investigation : the official journal of the International Society of Amyloidosis.

[3]  D. Borchelt,et al.  Axonal Transport of Mutant Superoxide Dismutase 1 and Focal Axonal Abnormalities in the Proximal Axons of Transgenic Mice , 1998, Neurobiology of Disease.

[4]  Robert H. Brown,et al.  Familial Amyotrophic Lateral Sclerosis-associated Mutations Decrease the Thermal Stability of Distinctly Metallated Species of Human Copper/Zinc Superoxide Dismutase* , 2002, The Journal of Biological Chemistry.

[5]  T. Lyons,et al.  The metal binding properties of the zinc site of yeast copper-zinc superoxide dismutase: implications for amyotrophic lateral sclerosis , 2000, JBIC Journal of Biological Inorganic Chemistry.

[6]  J T Finch,et al.  Amyloid fibers are water-filled nanotubes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[7]  A. Goldberg,et al.  Cellular Defenses against Unfolded Proteins A Cell Biologist Thinks about Neurodegenerative Diseases , 2001, Neuron.

[8]  J. Haines,et al.  Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis , 1993, Nature.

[9]  Michael A Hough,et al.  The structure of holo and metal-deficient wild-type human Cu, Zn superoxide dismutase and its relevance to familial amyotrophic lateral sclerosis. , 2003, Journal of molecular biology.

[10]  D. Bredesen,et al.  Altered Reactivity of Superoxide Dismutase in Familial Amyotrophic Lateral Sclerosis , 1996, Science.

[11]  E. Stadtman,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. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[12]  D. Borchelt,et al.  Fibrillar Inclusions and Motor Neuron Degeneration in Transgenic Mice Expressing Superoxide Dismutase 1 with a Disrupted Copper-Binding Site , 2002, Neurobiology of Disease.

[13]  J S Valentine,et al.  Copper(2+) binding to the surface residue cysteine 111 of His46Arg human copper-zinc superoxide dismutase, a familial amyotrophic lateral sclerosis mutant. , 2000, Biochemistry.

[14]  M. Beal,et al.  Motor neurons in Cu/Zn superoxide dismutase-deficient mice develop normally but exhibit enhanced cell death after axonal injury , 1996, Nature Genetics.

[15]  D. Borchelt,et al.  An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria , 1995, Neuron.

[16]  W. Hung,et al.  Intense Superoxide Dismutase‐1 Immunoreactivity in Intracytoplasmic Hyaline Inclusions of Familial Amyotrophic Lateral Sclerosis with Posterior Column Involvement , 1996, Journal of neuropathology and experimental neurology.

[17]  I. Fridovich,et al.  Amyotrophic lateral sclerosis: A proposed mechanism , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[18]  L. Bruijn,et al.  Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1. , 1998, Science.

[19]  R M Esnouf,et al.  Further additions to MolScript version 1.4, including reading and contouring of electron-density maps. , 1999, Acta crystallographica. Section D, Biological crystallography.

[20]  R. Read.  Improved Fourier Coefficients for Maps Using Phases from Partial Structures with Errors , 1986 .

[21]  L. Ellerby,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. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Nersissian,et al.  Loss of in Vitro Metal Ion Binding Specificity in Mutant Copper-Zinc Superoxide Dismutases Associated with Familial Amyotrophic Lateral Sclerosis* , 2000, The Journal of Biological Chemistry.

[23]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[24]  D. Price,et al.  Histological Evidence of Protein Aggregation in Mutant SOD1 Transgenic Mice and in Amyotrophic Lateral Sclerosis Neural Tissues , 2001, Neurobiology of Disease.

[25]  Robert H. Brown,et al.  Decreased Metallation and Activity in Subsets of Mutant Superoxide Dismutases Associated with Familial Amyotrophic Lateral Sclerosis* 210 , 2002, The Journal of Biological Chemistry.

[26]  K. Fischbeck,et al.  Toxic Proteins in Neurodegenerative Disease , 2002, Science.

[27]  J S Beckman,et al.  Induction of nitric oxide-dependent apoptosis in motor neurons by zinc-deficient superoxide dismutase. , 1999, Science.

[28]  Wendy Bruening,et al.  Up‐Regulation of Protein Chaperones Preserves Viability of Cells Expressing Toxic Cu/Zn‐Superoxide Dismutase Mutants Associated with Amyotrophic Lateral Sclerosis , 1999, Journal of neurochemistry.

[29]  D. Borchelt,et al.  ALS-Linked SOD1 Mutant G85R Mediates Damage to Astrocytes and Promotes Rapidly Progressive Disease with SOD1-Containing Inclusions , 1997, Neuron.

[30]  M. Pericak-Vance,et al.  Amyotrophic lateral sclerosis and structural defects in Cu,Zn superoxide dismutase. , 1993, Science.

[31]  C. Sander,et al.  Errors in protein structures , 1996, Nature.

[32]  M. Gurney,et al.  Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. , 1994, Science.

[33]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[34]  Matteo Pellegrini,et al.  Subunit asymmetry in the three‐dimensional structure of a human CuZnSOD mutant found in familial amyotrophic lateral sclerosis , 1998, Protein science : a publication of the Protein Society.

[35]  J. Chen,et al.  Reactions of nitric oxide, superoxide and peroxynitrite with superoxide dismutase in neurodegeneration. , 1994, Progress in brain research.

[36]  C R Kissinger,et al.  Rapid automated molecular replacement by evolutionary search. , 1999, Acta crystallographica. Section D, Biological crystallography.

[37]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[38]  K. Wilson,et al.  Efficient anisotropic refinement of macromolecular structures using FFT. , 1999, Acta crystallographica. Section D, Biological crystallography.

[39]  G N Murshudov,et al.  Use of TLS parameters to model anisotropic displacements in macromolecular refinement. , 2001, Acta crystallographica. Section D, Biological crystallography.

[40]  I. Fridovich.  Superoxide dismutases: an adaptation to a paramagnetic gas , 1989 .

[41]  G. Cohen.  Align : A program to superimpose protein coordinates, accounting for insertions and deletions , 1997 .

[42]  J. Morrison,et al.  Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  J. Richardson,et al.  Natural β-sheet proteins use negative design to avoid edge-to-edge aggregation , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[44]  D. Borchelt,et al.  High Molecular Weight Complexes of Mutant Superoxide Dismutase 1: Age-Dependent and Tissue-Specific Accumulation , 2002, Neurobiology of Disease.

[45]  Jeffrey Rothstein,et al.  Mutant SOD1 causes motor neuron disease independent of copper chaperone–mediated copper loading , 2002, Nature Neuroscience.

[46]  S H Appel,et al.  Natural history of amyotrophic lateral sclerosis in a database population. Validation of a scoring system and a model for survival prediction. , 1995, Brain : a journal of neurology.

[47]  Robert H. Brown,et al.  Superoxide Dismutase Activity, Oxidative Damage, and Mitochondrial Energy Metabolism in Familial and Sporadic Amyotrophic Lateral Sclerosis , 1993, Journal of neurochemistry.

[48]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[49]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[50]  M. Gurney,et al.  Formation of high molecular weight complexes of mutant Cu, Zn-superoxide dismutase in a mouse model for familial amyotrophic lateral sclerosis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[51]  A. F. Soleng,et al.  A thorny question: how does activity maintain dendritic spines? , 1999, Nature Neuroscience.

[52]  Jorge Navaza,et al.  [33] AMoRe: An automated molecular replacement program package. , 1997, Methods in enzymology.

[53]  T. Yeates,et al.  Arrangement of subunits and ordering of β-strands in an amyloid sheet , 2002, Nature Structural Biology.

[54]  J. Thompson,et al.  Decreased Zinc Affinity of Amyotrophic Lateral Sclerosis‐Associated Superoxide Dismutase Mutants Leads to Enhanced Catalysis of Tyrosine Nitration by Peroxynitrite , 1997, Journal of neurochemistry.