Dimerization, Oligomerization, and Aggregation of Human Amyotrophic Lateral Sclerosis Copper/Zinc Superoxide Dismutase 1 Protein Mutant Forms in Live Cells

Kim, Jiho; Lee, Honggun; Lee, Juhyun; Kwon, Doyoon; Genovesio, Auguste; Fenistein, Denis; Ogier, Arnaud; Brondani, Vincent; Grailhe, Régis

doi:10.1074/jbc.m113.542613

Cited by 47 publications

(52 citation statements)

References 25 publications

(13 reference statements)

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“…7–9 We first tested whether SOD1 G93A also forms aggregates in the intestinal epithelial cells. The human intestinal epithelial cells (HCT116) were transfected with fluorescent protein tagged human wild type SOD1-GFP or human ALS mutation SOD1 G93A -GFP.…”

Section: Resultsmentioning

confidence: 99%

“…6 Intestinal epithelial cells are consistently exposed to bacteria, a process which plays a key role in development, renewal, and immunity. 7–9 Frequent microbial challenges to epithelial cells trigger discrete signaling pathways, promoting molecular changes, such as the secretion of cytokines and chemokines, and alterations to molecules displayed at the epithelial cell surface. Intestinal homeostasis and microbiome play essential roles in neurological diseases, such as autism and Parkinson’s disease.…”

Section: Introductionmentioning

confidence: 99%

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Target Intestinal Microbiota to Alleviate Disease Progression in Amyotrophic Lateral Sclerosis

Zhang

et al. 2017

Clinical Therapeutics

214

175

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Purpose Emerging evidence has demonstrated that gut microbiome plays essential roles in the pathogenesis of human diseases in distant organs. Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive loss of motor neurons. Treatment with the only FDA approved drug, Riluzole, extends patient life span only for a few months. Thus, there is an urgent need to develop novel interventions for alleviating the disease progression and improving the quality of life for ALS patients. Here we present evidence that intestinal dysfunction and dysbiosis may actively contribute to ALS pathophysiology. Methods We used G93A transgenic mice as a model of human ALS. The G93A mice show abnormal intestinal microbiome and damaged tight junctions, which occur before ALS disease onset. Feeding of the G93A mice with butyrate, a natural bacterial product. Results Feeding of the G93A mice with butyrate, restores the intestinal microbial homeostasis, improves the gut integrity, and prolongs the life span of the ALS mice. Paneth cells are specialized intestinal epithelial cells that regulate the host-bacterial interactions. At the cellular level, we show that abnormal Paneth cells were significantly decreased in the ALS mice treated by butyrate. Butyrate treatment decreases aggregation of the G93A-SOD1 mutated protein in both ALS mice and cultured human intestinal epithelial cells. Implications Our study highlights the complex role of gut microbiome and intestinal epithelium in the progression of ALS and presents butyrate as a potential therapeutic reagent to restore ALS dysbiosis.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Target Intestinal Microbiota to Alleviate Disease Progression in Amyotrophic Lateral Sclerosis

Zhang

et al. 2017

Clinical Therapeutics

214

175

View full text Add to dashboard Cite

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“…(8–12) Aggregation results primarily from mutations in SOD1, and over 100 mutant forms of SOD1 have been linked to ALS. (8,13–15)…”

Section: Introductionmentioning

confidence: 99%

Cu,Zn-Superoxide Dismutase without Zn Is Folded but Catalytically Inactive

Nedd

Redler

Proctor

et al. 2014

Journal of Molecular Biology

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Amyotrophic Lateral Sclerosis has been linked to the gain of aberrant function of superoxide dismutase, Cu,Zn-SOD1 upon protein misfolding. The mechanism of SOD1 misfolding is thought to involve mutations leading to the loss of Zn, followed by protein unfolding, and aggregation. We show that the removal of Zn from SOD1 may not lead to an immediate unfolding, but immediately deactivates the enzyme through a combination of subtle structural and electronic effects. Using Quantum Mechanics/Discrete Molecular Dynamics, we showed that Zn-less wild type SOD1 and its D124N mutant that does not bind Zn both have at least metastable folded states. In those states, the reduction potential of Cu increases, leading to the presence of detectable amounts of Cu(I) instead of Cu(II) in the active site, as confirmed experimentally. The Cu(I) protein cannot participate in the catalytic Cu(I) – Cu(II) cycle. However, even without the full reduction to Cu(I), the Cu site in the Zn-less variants of SOD1 is shown to be catalytically incompetent: unable to bind superoxide in a way comparable to the wild type SOD1. The changes are more radical and different in the D124N Zn-less mutant than in the Zn-less wild type SOD1, suggesting D124N being perhaps not the most adequate model for Zn-less SOD1. Overall, Zn in SOD1 appears to be influencing the Cu site directly by adjusting its reduction potential and geometry. Thus, the role of Zn in SOD1 is not just structural, as was previously thought; it is a vital part of the catalytic machinery.

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“…It is therefore unclear whether the decrease in enzymatic activity of mutant SOD1 is a deciding factor for its neurotoxic action . On the other hand, it is suggested that a monomeric SOD1 is prone to be misfolded and/or aggregated forms and they trigger various neuronal dysfunctions and degeneration . Among these mutations, we chose 2 well‐investigated mutations, G85R and G93A, and studied their effects on SOD1 dimerization, assembly of the NanoBit system and luciferase activity in living cells.…”

Section: Resultsmentioning

confidence: 99%

“…3,4 These mutations are suggested to decrease their conformational stability and trigger their monomeric form. 16,[18][19][20] In addition, these mutations reduce their enzymatic activities in varying proportions but do not abolish them completely. 16 It is therefore unclear whether the decrease in enzymatic activity of mutant SOD1 is a deciding factor for its neurotoxic action.…”

Section: Resultsmentioning

confidence: 99%

SOD1 dimerization monitoring using a novel split NanoLuc, NanoBit

Oh‐hashi

Hirata

Kiuchi

2016

Cell Biochemistry & Function

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In the present study, we applied a highly sensitive NanoLuc-based technology to understand the status of superoxide dismutase 1 (SOD1) within mammalian cells. Two fragments of NanoLuc (NanoBit), large N-terminal and small C-terminal regions, were fused with wild-type (wt) and mutant human SOD1 (hSOD1) genes and transfected into cells. Luciferase activity through NanoBit assembly was only detected in NanoBit-tagged wtSOD1-expressing cells. Furthermore, the developed NanoLuc system was used to investigate the role of protein-protein interactions in the pathogenesis of amyotrophic lateral sclerosis (ALS). In addition to SOD1, we also applied this NanoBit system for detecting the dimerization of wild-type, M337V-mutated human TAR-binding protein 43 kDa (hTDP43) and its cleaved C-terminal fragment (TDP25 ) as well as their interactions with SOD1. Luciferase activities of NanoBit-tagged mutant SOD1, TDP43, or TDP25 were negligible. Finally, we found that a zinc chelator partially reduced the luciferase activity of NanoBit-wtSOD1. Collectively, these results show that the present assay is sensitive and convenient to appreciate ALS and to develop useful agents for the modulation of SOD1 conformation.

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Dimerization, Oligomerization, and Aggregation of Human Amyotrophic Lateral Sclerosis Copper/Zinc Superoxide Dismutase 1 Protein Mutant Forms in Live Cells

Cited by 47 publications

References 25 publications

Target Intestinal Microbiota to Alleviate Disease Progression in Amyotrophic Lateral Sclerosis

Target Intestinal Microbiota to Alleviate Disease Progression in Amyotrophic Lateral Sclerosis

Cu,Zn-Superoxide Dismutase without Zn Is Folded but Catalytically Inactive

SOD1 dimerization monitoring using a novel split NanoLuc, NanoBit

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