Structural and biophysical properties of metal-free pathogenic SOD1 mutants A4V and G93A

Galaleldeen, Ahmad; Strange, R.W.; Whitson, Lisa J.; Antonyuk, S.V.; Narayana, Narendra; Taylor, Alexander B.; Schuermann, Jonathan P.; Holloway, S.; Hasnain, S.S.; Hart, P. John

doi:10.1016/j.abb.2009.09.020

Cited by 75 publications

(82 citation statements)

References 64 publications

(82 reference statements)

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“…We present here further characterization of the C4F6 antibody, which is known to react specifically with mutant human SOD1 (hSOD1) proteins (20,24). The antibody was generated against the apo (metal-free) form of hSOD1 G93A , which exhibits a disordered conformation (25). Immunization with recombinant hSOD1 G93A protein increases the life span of hSOD1 G37R mice while simultaneously decreasing C4F6 detection of mutant hSOD1 protein, suggesting that C4F6 recognizes toxic mutant SOD1 (24).…”

mentioning

confidence: 99%

Localization of a toxic form of superoxide dismutase 1 protein to pathologically affected tissues in familial ALS

Brotherton

Cooper

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Mutations in the gene encoding superoxide dismutase 1 (SOD1) account for about 20% of the cases of familial amyotrophic lateral sclerosis (fALS). It is not known how the mutant protein causes disease, or why only a subset of cell types (motor neurons) are targeted. The aggregation and misfolding of mutant SOD1 are implicated in disease pathogenesis in both animal models and humans. We used a monoclonal antibody, C4F6, which specifically reacts with mutant and/or "misfolded" SOD1, to investigate the regional distribution of mutant SOD1 protein in rodent and human tissues. C4F6 reacted only with mutant SOD1 and showed remarkable selectivity for disease-affected tissues and cells. Tissue not affected by disease but containing high levels of mutant protein (sensory neurons) did not stain with C4F6. Additionally, C4F6 intensely stained some motor neurons while leaving adjacent motor neurons unstained. Although C4F6 was generated against the G93A SOD1 mutant, it also recognized other SOD1 mutants. In human autopsy tissues from patients carrying SOD1 mutations, C4F6 identified skein-like intracellular inclusions in motor neurons, similar to those seen in rodents, and again stained only a subset of motor neurons. In spinal cords from patients with sporadic ALS, other neurodegenerative diseases, and normal controls, C4F6-immunoreactive inclusions were not detected, but the antibody did reveal diffuse immunostaining of some spinal motor neurons. The ability of C4F6 to differentiate pathologically affected tissue in mutant SOD1 ALS rodent models and humans, specifically motor neuron populations, suggests that this antibody may recognize a "toxic" form of the mutant SOD1 protein.aggregate | misfold | soluble A myotrophic lateral sclerosis (ALS) is a progressive, adultonset motor neuron disease with an incidence of one to two persons per 100,000 (1, 2). ALS typically causes death within 3-5 y of diagnosis; there is no cure and treatment options are very limited (3-5). Ten percent of ALS cases are familial (fALS), with 20% of fALS being linked to mutations in the homodimeric protein Cu/Zn superoxide dismutase 1 (SOD1) (6-8). In animal models and in humans carrying SOD1 mutations, mutant SOD1 is "toxic" only to motor neurons, and only after it has been present for an extended period (aging), leading to the important questions of how mutant SOD1 exerts its toxicity and why motor neurons are particularly sensitive to the pathogenic process.Mutant SOD1 has a high propensity to undergo conformational changes and aggregation when stressed in vitro, and SOD1 aggregates and misfolded aggregate precursors have been localized specifically to pathologically affected tissues in animal models of disease, suggesting that SOD1 conformational changes may contribute to the toxicity of the mutant protein (9)(10)(11)(12)(13)(14). It has been further demonstrated that conformational changes in wildtype SOD1 can be induced through oxidation and/or metal depletion and that these conformers of the wild-type protein are also toxic in cellular models...

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confidence: 99%

Localization of a toxic form of superoxide dismutase 1 protein to pathologically affected tissues in familial ALS

Brotherton

Cooper

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…It has been reported that SOD aggregation is almost entirely prevented by addition of Cu 2+ and Zn 2+ ions that stabilize the native-like conformation [16]. Inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis showed the presence of Cu 2+ and Zn 2+ ions in rAhSOD in a ratio of 0.2502 and 0.2612 μg/mg protein, respectively.…”

Section: Metal Identification In Rahsodmentioning

confidence: 99%

“…In medicine, recombinant SODs may be useful for the treatment of a wide variety of disorders where oxidative stress is involved such as cancer, inflammation, obesity, diabetes, amyotrophic lateral sclerosis, and Alzheimer's disease [12][13][14][15][16][17]. Therefore, SOD enzymes are actively investigated as potential therapeutic agents in diseases related to oxidative stress and for their role in moderating the ageing process [18].…”

Section: Introductionmentioning

confidence: 99%

Biochemical and Molecular Characterization of a Novel Cu/Zn Superoxide Dismutase from Amaranthus hypochondriacus L.: an Intrinsically Disordered Protein

Montero-Morán

Sampedro

Saab‐Rincón³

et al. 2015

Appl Biochem Biotechnol

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A novel Cu/ZnSOD from Amaranthus hypochondriacus was cloned, expressed, and characterized. Nucleotide sequence analysis showed an open reading frame (ORF) of 456 bp, which was predicted to encode a 15.6-kDa molecular weight protein with a pI of 5.4. Structural analysis showed highly conserved amino acid residues involved in Cu/Zn binding. Recombinant amaranth superoxide dismutase (rAhSOD) displayed more than 50 % of catalytic activity after incubation at 100 °C for 30 min. In silico analysis of Amaranthus hypochondriacus SOD (AhSOD) amino acid sequence for globularity and disorder suggested that this protein is mainly disordered; this was confirmed by circular dichroism, which showed the lack of secondary structure. Intrinsic fluorescence studies showed that rAhSOD undergoes conformational changes in two steps by the presence of Cu/Zn, which indicates the presence of two binding sites displaying different affinities for metals ions. Our results show that AhSOD could be classified as an intrinsically disordered protein (IDP) that is folded when metals are bound and with high thermal stability.

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“…First, the mutation could induce structural perturbations that result in alterations of the chemical environment of Cys-111. In this regard, it has been shown that mutation G93A leads to noticeable changes in the structure and dynamics of hSOD1 (53,54). In addition, the mutation may render Cys-111 more sensitive to oxidation (47,48), which would make Cys-111 unreactive.…”

Section: Detection Of Thiol/disulfide Exchanges From the Length Signamentioning

confidence: 99%

Altered Thiol Chemistry in Human Amyotrophic Lateral Sclerosis-linked Mutants of Superoxide Dismutase 1

Solsona

Kahn

Badilla

et al. 2014

Journal of Biological Chemistry

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Background: Lou Gehrig's disease is accompanied by misfolding and aggregation of proteins. Results: The intramolecular reactivity of cysteines of superoxide dismutase 1 results in new disulfide bonds in unusual positions. Conclusion: Thiol/disulfide exchange reactions are altered in mutated proteins. Significance: Intramolecular reorganization of disulfides may be one of the mechanisms of protein misfolding related to neurodegenerative diseases.

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Structural and biophysical properties of metal-free pathogenic SOD1 mutants A4V and G93A

Cited by 75 publications

References 64 publications

Localization of a toxic form of superoxide dismutase 1 protein to pathologically affected tissues in familial ALS

Localization of a toxic form of superoxide dismutase 1 protein to pathologically affected tissues in familial ALS

Biochemical and Molecular Characterization of a Novel Cu/Zn Superoxide Dismutase from Amaranthus hypochondriacus L.: an Intrinsically Disordered Protein

Altered Thiol Chemistry in Human Amyotrophic Lateral Sclerosis-linked Mutants of Superoxide Dismutase 1

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