Details of a Researcher - Furukawa, Yoshiaki

Papers - Furukawa, Yoshiaki

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Cysteine residues in Cu,Zn-superoxide dismutase are essential to toxicity in Caenorhabditis elegans model of amyotrophic lateral sclerosis

Ogawa, Mariko, Shidara, Hisashi, Oka, Kotaro, Kurosawa, Masaru, Nukina, Nobuyuki, Furukawa, Yoshiaki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 463 ( 4 ) 1196 - 1202 2015.08

Research paper (scientific journal), Joint Work, Accepted, ISSN 0006-291X

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Conformational Disorder of the Most Immature Cu, Zn-Superoxide Dismutase Leading to Amyotrophic Lateral Sclerosis

Furukawa, Yoshiaki, Anzai, Itsuki, Akiyama, Shuji, Imai, Mizue, Cruz, Fatima Joy C., Saio, Tomohide, Nagasawa, Kenichi, Nomura, Takao, Ishimori, Koichiro

JOURNAL OF BIOLOGICAL CHEMISTRY 291 ( 8 ) 4144 - 4155 2016.02

Research paper (scientific journal), Joint Work, Accepted, ISSN 0021-9258

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A molecular mechanism realizing sequence-specific recognition of nucleic acids by TDP-43

Furukawa, Yoshiaki, Suzuki, Yoh, Fukuoka, Mami, Nagasawa, Kenichi, Nakagome, Kenta, Shimizu, Hideaki, Mukaiyama, Atsushi, Akiyama, Shuji

SCIENTIFIC REPORTS 6 4144 - 4155 2016.02

Research paper (scientific journal), Joint Work, Accepted, ISSN 2045-2322

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Oxidative denaturation of Cu/Zn-superoxide dismutase associated with neurodegenerative diseases

Yoshida M., Muraki N., Tajiri M., Hengphasatporn K., Sue K., Kubo T., Akashi S., Shigeta Y., Kambe T., Furukawa Y.

Protein Science 34 ( 11 ) 2025.11

ISSN 09618368

Misfolding of mutant Cu/Zn-superoxide dismutase (SOD1) is a well-established pathological feature of familial amyotrophic lateral sclerosis (ALS). While amino acid substitutions in mutant SOD1 destabilize its structure and promote misfolding, oxidation has also been implicated in the pathological alterations of wild-type SOD1, particularly in neurodegenerative diseases including sporadic ALS. However, the impact of oxidation on SOD1 folding remains to be fully elucidated. Here, we demonstrate that Cys111 is primarily oxidized to sulfonic acid upon exposure of apo-SOD1 to hydrogen peroxide, as confirmed by the quantitation of thiol groups and mass spectrometry. Molecular dynamics simulations showed that sulfonylation of Cys111 disrupts the dimer interface and promotes monomerization. This monomeric form then facilitates the subsequent oxidation of buried Cys6, leading to structural disruption, as evidenced by circular dichroism spectroscopy and loss of thiol groups. SOD1 denaturation triggered by Cys111 oxidation became evident when zinc binding was impaired due to pathological mutations and/or under zinc-deficient conditions. Given that increased oxidative stress is frequently associated with many neurodegenerative diseases, modulating Cys111 oxidation may offer a potential strategy for maintaining SOD1 structural stability and preventing its pathological misfolding.

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Cu/Zn-superoxide dismutase naturally fused with a β-propeller lactonase in Deinococcus radiodurans.

Yoshiaki Furukawa, Masamichi Megata, Atsuko Shintani, Kaori Sue, Tomohiro Morohoshi, Masato Akutsu, Norifumi Muraki

The Journal of biological chemistry 301 ( 8 ) 110499 - 110499 2025.08

ISSN 00219258

Cu/Zn-superoxide dismutase (Cu/Zn-SOD) is an antioxidant enzyme widely present across species; however, the structural diversity and physiological roles of Cu/Zn-SOD are yet to be fully uncovered. Here, we show a unique type of Cu/Zn-SOD from Deinococcus radiodurans (DrSOD) with an additional β-propeller domain. Our structural analysis on DrSOD revealed a typical bacterial Cu/Zn-SOD domain, binding both a copper and zinc ion, alongside a six-bladed β-propeller domain coordinating a calcium ion. DrSOD was indeed expressed in D. radiodurans, but its deletion did not lead to any noticeable changes in resistance to DNA-damaging stresses, a characteristic trait of D. radiodurans. Despite this, the Cu/Zn-SOD domain retained superoxide dismutase activity, and the β-propeller domain was found to exhibit a lactonase activity specifically for hydrolyzing 2-coumaranone. Taken together, while the precise physiological role of DrSOD needs to be further investigated, our findings here reveal a unique multi-functional enzyme architecture, expanding the known structural diversity of Cu/Zn-SODs.

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Metal-Responsive Up-Regulation of Bifunctional Disulfides for Suppressing Protein Misfolding and Promoting Oxidative Folding.

Keita Mori, Tsubura Kuramochi, Motonori Matsusaki, Yuki Hashiguchi, Masaki Okumura, Tomohide Saio, Yoshiaki Furukawa, Kenta Arai, Takahiro Muraoka

Angewandte Chemie (International ed. in English) 64 ( 36 ) e202502187 2025

ISSN 14337851

The stress-responsive up-regulation process is a sophisticated biological response to maintain cellular homeostasis. In intracellular anti-oxidant systems, the expression level of oxidoreductases is up-regulated under oxidative stress, mitigating oxidative damage on biomolecules and enhancing protein folding capacity. Herein, inspired by the biological system, we developed a synthetic folding promotor whose reactivity is up-regulated under stress conditions. We conjugated two metal-binding 1,4,7,11-tetraazacyclotetradecane (cyclam) ligands and a redox-active disulfide to obtain cyclam-SS, whose reactivity can be enhanced under metal-induced stress. Metal coordination increased the redox potential of cyclam-SS, activating it as an oxidant. While CuII ions severely hampered the oxidative folding of substrate polypeptides, cyclam-SS exhibited bifunctional folding-promoting properties, i) suppressing CuII-mediated misfolding and aggregation, and ii) harnessing CuII to enhance oxidative folding. Cyclam-SS was also useful for disulfide-bond formation to promote oxidative folding of pharmaceutical and pathological proteins, as demonstrated with proinsulin and superoxide dismutase 1 (SOD1). Furthermore, cyclam-SS protected cultured cells from copper-induced stress. Thus, we demonstrated the induction of the stress-responsive up-regulation process by a bifunctional folding promotor controlling the folding status of biologically important proteins under metal-induced stress. The strategy of "stress-responsive up-regulation" could aid the development of novel synthetic materials for treating intracellular stress and related disorders.

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Disulfide-mediated oligomerization of mutant Cu/Zn-superoxide dismutase associated with canine degenerative myelopathy.

Yuki Shino, Norifumi Muraki, Yui Kobatake, Hiroaki Kamishina, Ryuichi Kato, Yoshiaki Furukawa

Protein science : a publication of the Protein Society 33 ( 12 ) e5210 2024.12

ISSN 09618368

A homozygous E40K mutation in the gene coding canine Cu/Zn-superoxide dismutase (cSOD1) causes degenerative myelopathy (DM) in dogs. A pathological hallmark of DM with the cSOD1 mutation is the aggregation of mutant cSOD1 proteins in neurons. The amino acid substitution E40K disrupts a salt bridge between Glu40 and Lys91 and is considered to destabilize the native state of cSOD1; however, the mechanism by which mutant cSOD1 aggregates remains unclear. Here, we show that mutant cSOD1 losing a copper and zinc ion forms oligomers crosslinked via disulfide bonds. The E40K substitution was found to result in the increased solvent exposure of the Cys7 side chain, which then attacked the disulfide bond (Cys57-Cys146) in cSOD1 to form disulfide-linked oligomers. We also successfully prevented the Cys7 exposure and thus the oligomerization of mutant cSOD1 by a fragment antibody that specifically recognizes the region around the mutation site. The fragment antibody covered the β-plug region, reinforcing the interactions compromised by the E40K substitution and thus contributing to the maintenance of the structural integrity of the β-barrel core of cSOD1. Taken together, we propose that the Cys7 exposure in cSOD1 upon the salt bridge disruption plays a central role in the aggregation mechanism of DM-associated mutant cSOD1.

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Engineering a monobody specific to monomeric Cu/Zn-superoxide dismutase associated with amyotrophic lateral sclerosis.

Hiroshi Amesaka, Mizuho Hara, Yuki Sakai, Atsuko Shintani, Kaori Sue, Tomoyuki Yamanaka, Shun-Ichi Tanaka, Yoshiaki Furukawa

Protein science : a publication of the Protein Society 33 ( 4 ) e4961 2024.04

ISSN 09618368

Misfolding of mutant Cu/Zn-superoxide dismutase (SOD1) has been implicated in familial form of amyotrophic lateral sclerosis (ALS). A natively folded SOD1 forms a tight homodimer, and the dimer dissociation has been proposed to trigger the oligomerization/aggregation of SOD1. Besides increasing demand for probes allowing the detection of monomerized forms of SOD1 in various applications, the development of probes has been limited to conventional antibodies. Here, we have developed Mb(S4) monobody, a small synthetic binding protein based on the fibronectin type III scaffold, that recognizes a monomeric but not dimeric form of SOD1 by performing combinatorial library selections using phage and yeast-surface display methods. Although Mb(S4) was characterized by its excellent selectivity to the monomeric conformation of SOD1, the monomeric SOD1/Mb(S4) complex was not so stable (apparent Kd ~ μM) as to be detected in conventional pull-down experiments. Instead, the complex of Mb(S4) with monomeric but not dimeric SOD1 was successfully trapped by proximity-enabled chemical crosslinking even when reacted in the cell lysates. We thus anticipate that Mb(S4) binding followed by chemical crosslinking would be a useful strategy for in vitro and also ex vivo detection of the monomeric SOD1 proteins.

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Characterization of a novel cysteine-less Cu/Zn-superoxide dismutase in Paenibacillus lautus missing a conserved disulfide bond.

Yoshiaki Furukawa, Atsuko Shintani, Shuhei Narikiyo, Kaori Sue, Masato Akutsu, Norifumi Muraki

The Journal of biological chemistry 299 ( 8 ) 105040 - 105040 2023.08

ISSN 00219258

Cu/Zn-superoxide dismutase (CuZnSOD) is an enzyme that binds a copper and zinc ion and also forms an intramolecular disulfide bond. Together with the copper ion as the active site, the disulfide bond is completely conserved among these proteins; indeed, the disulfide bond plays critical roles in maintaining the catalytically competent conformation of CuZnSOD. Here, we found that a CuZnSOD protein in Paenibacillus lautus (PaSOD) has no Cys residue but exhibits a significant level of enzyme activity. The crystal structure of PaSOD revealed hydrophobic and hydrogen-bonding interactions in substitution for the disulfide bond of the other CuZnSOD proteins. Also notably, we determined that PaSOD forms a homodimer through an additional domain with a novel fold at the N terminus. While the advantages of lacking Cys residues and adopting a novel dimer configuration remain obscure, PaSOD does not require a disulfide-introducing/correcting system for maturation and could also avoid misfolding caused by aberrant thiol oxidations under an oxidative environment.

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Intrinsic structural vulnerability in the hydrophobic core induces species-specific aggregation of canine SOD1 with degenerative myelopathy-linked E40K mutation.

Kei Hashimoto, Seiji Watanabe, Masato Akutsu, Norifumi Muraki, Hiroaki Kamishina, Yoshiaki Furukawa, Koji Yamanaka

The Journal of biological chemistry 299 ( 6 ) 104798 - 104798 2023.06

ISSN 00219258

Canine degenerative myelopathy (DM), a fatal neurodegenerative disease in dogs, shares clinical and genetic features with amyotrophic lateral sclerosis (ALS), a human motor neuron disease. Mutations in the SOD1 gene encoding Cu/Zn superoxide dismutase (SOD1) cause canine DM and a subset of inherited human ALS. The most frequent DM causative mutation is homozygous E40K mutation which induces the aggregation of canine SOD1 but not of human SOD1. However, the mechanism through which canine E40K mutation induces species-specific aggregation of SOD1 remains unknown. By screening human/canine chimeric SOD1s, we identified that the humanized mutation of the 117th residue (M117L), encoded by exon 4, significantly reduced aggregation propensity of canine SOD1E40K. Conversely, introducing a mutation of leucine 117 to methionine, a residue homologous to canine, promoted E40K-dependent aggregation in human SOD1. M117L mutation improved protein stability and reduced cytotoxicity of canine SOD1E40K. Furthermore, crystal structural analysis of canine SOD1 proteins revealed that M117L increased the packing within the hydrophobic core of the β-barrel structure, contributing to the increased protein stability. Our findings indicate that the structural vulnerability derived intrinsically from Met 117 in the hydrophobic core of the β-barrel structure induces E40K-dependent species-specific aggregation in canine SOD1.

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Zinc-mediated interaction of copper chaperones through their heavy-metal associated domains

Furukawa Y., Matsumoto K., Nakagome K., Shintani A., Sue K.

Journal of Trace Elements in Medicine and Biology 75 2023.01

ISSN 0946672X

Background: A copper chaperone CCS is a multi-domain protein that supplies a copper ion to Cu/Zn-superoxide dismutase (SOD1). Among the domains of CCS, the N-terminal domain (CCSdI) belongs to a heavy metal-associated (HMA) domain, in which a Cys-x-x-Cys (CxxC) motif binds a heavy metal ion. It has hence been expected that the HMA domain in CCS has a role in the metal trafficking; however, the CxxC motif in the domain is dispensable for supplying a copper ion to SOD1, leaving an open question on roles of CCSdI in CCS. Methods: To evaluate protein-protein interactions of CCS through CCSdI, yeast two-hybrid assay, a pull-down assay using recombinant proteins, and the analysis with fluorescence resonance energy transfer were performed. Results: We found that CCS specifically interacted with another copper chaperone HAH1, a HMA domain protein, through CCSdI. The interaction between CCSdI and HAH1 was not involved in the copper supply from CCS to SOD1 but was mediated by a zinc ion ligated with Cys residues of the CxxC motifs in CCSdI and HAH1. Conclusion: While physiological significance of the interaction between copper chaperones awaits further investigation, we propose that CCSdI would have a role in the metal-mediated interaction with other proteins including heterologous copper chaperones.

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Quantum-dot-labeled synuclein seed assay identifies drugs modulating the experimental prion-like transmission.

Yukio Imamura, Ayami Okuzumi, Saki Yoshinaga, Akiko Hiyama, Yoshiaki Furukawa, Tomohiro Miyasaka, Nobutaka Hattori, Nobuyuki Nukina

Communications biology 5 ( 1 ) 636 - 636 2022.12

Synucleinopathies are neurodegenerative disorders including Parkinson disease (PD), dementia with Lewy body (DLB), and multiple system atrophy (MSA) that involve deposits of the protein alpha-synuclein (α-syn) in the brain. The inoculation of α-syn aggregates derived from synucleinopathy or preformed fibrils (PFF) formed in vitro induces misfolding and deposition of endogenous α-syn. This is referred to as prion-like transmission, and the mechanism is still unknown. In this study, we label α-syn PFF with quantum dots and visualize their movement directly in acute slices of brain tissue inoculated with α-syn PFF seeds. Using this system, we find that the trafficking of α-syn seeds is dependent on fast axonal transport and the seed spreading is dependent on endocytosis and neuronal activity. We also observe pharmacological effects on α-syn seed spreading; clinically available drugs including riluzole are effective in reducing the spread of α-syn seeds and this effect is also observed in vivo. Our quantum-dot-labeled α-syn seed assay system combined with in vivo transmission experiment reveals an early phase of transmission, in which uptake and spreading of seeds occur depending on neuronal activity, and a later phase, in which seeds induce the propagation of endogenous misfolded α-syn.

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FUS regulates RAN translation through modulating the G-quadruplex structure of GGGGCC repeat RNA inC9orf72-linked ALS/FTD

Yuzo Fujino, Morio Ueyama, Taro Ishiguro, Daisaku Ozawa, Toshihiko Sugiki, Hayato Ito, Asako Murata, Akira Ishiguro, Tania F. Gendron, Kohji Mori, Eiichi Tokuda, Tomoya Taminato, Takuya Konno, Akihide Koyama, Yuya Kawabe, Toshihide Takeuchi, Yoshiaki Furukawa, Toshimichi Fujiwara, Manabu Ikeda, Toshiki Mizuno, Hideki Mochizuki, Hidehiro Mizusawa, Keiji Wada, Kinya Ishikawa, Osamu Onodera, Kazuhiko Nakatani, Hideki Taguchi, Leonard Petrucelli, Yoshitaka Nagai

(eLife Sciences Publications, Ltd) 2022.11

Abnormal expansions of GGGGCC repeat sequence in the noncoding region of the C9orf72 gene is the most common cause of familial amyotrophic lateral sclerosis and frontotemporal dementia (C9-ALS/FTD). The expanded repeat sequence is translated into dipeptide repeat proteins (DPRs) by noncanonical repeat-associated non-AUG (RAN) translation. Since DPRs play central roles in the pathogenesis of C9-ALS/FTD, we here investigate the regulatory mechanisms of RAN translation, focusing on the effects of RNA-binding proteins (RBPs) targeting GGGGCC repeat RNAs. Using C9-ALS/FTD model flies, we demonstrated that the ALS/FTD-linked RBP FUS suppresses RAN translation and neurodegeneration in an RNA-binding activity-dependent manner. Moreover, we found that FUS directly binds to and modulates the G-quadruplex structure of GGGGCC repeat RNA as an RNA chaperone, resulting in the suppression of RAN translation in vitro. These results reveal a previously unrecognized regulatory mechanism of RAN translation by G-quadruplex-targeting RBPs, providing therapeutic insights for C9-ALS/FTD and other repeat expansion diseases.

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A rapid and simple electrochemical detection of the free drug concentration in human serum using boron-doped diamond electrodes

Moriyama H., Ogata G., Nashimoto H., Sawamura S., Furukawa Y., Hibino H., Kusuhara H., Einaga Y.

Analyst 147 ( 20 ) 4442 - 4449 2022.09

ISSN 00032654

Monitoring drug concentration in blood and reflecting this in the dosage are crucial for safe and effective drug treatment. Most drug assays are based on total concentrations of bound and unbound proteins in the serum, although only the unbound concentration causes beneficial and adverse events. Monitoring the unbound concentration alone is expected to provide a means for further optimisation of drug treatment. However, unbound concentration monitoring has not been routinely used for drug treatment due to the long analysis time and the high cost of conventional methods. Here, we have developed a rapid electrochemical method to determine the unbound concentration in ultrafiltered human serum using boron-doped diamond (BDD) electrodes. When the anticancer drug doxorubicin was used as the test drug, the catalytic doxorubicin-mediated reduction of dissolved oxygen provided a sensitive electrochemical signal, with a detection limit of 0.14 nM. In contrast, the sensitivity of glassy carbon (GC) was inferior under the same conditions due to interference from the dissolved oxygen reduction current. The signal background ratio (S/B) of BDD and GC was 11.5 (10 nM doxorubicin) and 1.1 (50 nM), respectively. The results show that a fast measurement time within ten seconds is possible in the clinical concentration range. Additionally, in the ultrafiltered human serum, the obtained values of unbound doxorubicin concentration showed good agreement with those quantified by conventional liquid chromatography-mass spectrometry. This approach has the potential for application in clinical settings where rapid and simple analysis methods would be beneficial.

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Novel oxindole compounds inhibit the aggregation of amyloidogenic proteins associated with neurodegenerative diseases.

Shintaro Kimura, Hiroaki Kamishina, Yoko Hirata, Kyoji Furuta, Yoshiaki Furukawa, Osamu Yamato, Sadatoshi Maeda, Yuji O Kamatari

Biochimica et biophysica acta. General subjects (Biochimica et Biophysica Acta - General Subjects) 1866 ( 5 ) 130114 - 130114 2022.05

ISSN 03044165

Amyloidogenic proteins form aggregates in cells, thereby leading to neurodegenerative disorders, including Alzheimer's and prion's disease, amyotrophic lateral sclerosis (ALS) in humans, and degenerative myelopathy (DM) and cognitive dysfunction in dogs. Hence, many small-molecule compounds have been screened to examine their inhibitory effects on amyloidogenic protein aggregation. However, no effective drug suitable for transition to clinical use has been found. Here we examined several novel oxindole compounds (GIF compounds) for their inhibitory effects on aggregate formation of the canine mutant superoxide dismutase 1 (cSOD1 E40K), a causative mutation resulting in DM, using Thioflavin-T fluorescence. Most GIF compounds inhibited the aggregation of cSOD1 E40K. Among the compounds, GIF-0854-r and GIF-0890-r were most effective. Their inhibitory effects were also observed in cSOD1 E40K-transfected cells. Additionally, GIF-0890-r effectively inhibited the aggregate formation of human SOD1 G93A, a causative mutation of ALS. GIF-0827-r and GIF-0856-r also effectively inhibited aggregate formation of human prion protein (hPrP). Subsequently, the correlation between their inhibitory effects on cSOD1 and hPrP aggregation was shown, indicating GIF compounds inhibited the aggregate formation of multiple amyloidogenic proteins. Conclusively, the novel oxindole compounds (GIF-0827-r, GIF-0854-r, GIF-0856-r, and GIF-0890-r) are proposed as useful therapeutic candidates for amyloidogenic neurodegenerative disorders.

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Metal distribution in Cu/Zn-superoxide dismutase revealed by native mass spectrometry.

Michiko Tajiri, Hiroto Aoki, Atsuko Shintani, Kaori Sue, Satoko Akashi, Yoshiaki Furukawa

Free radical biology & medicine (Free Radical Biology and Medicine) 183 60 - 68 2022.04

ISSN 08915849

Cu/Zn-superoxide dismutase (SOD1) is a homodimer with two identical subunits, each of which binds a copper and zinc ion in the native state. In contrast to such a text book case, SOD1 proteins purified in vitro or even in vivo have been often reported to bind a non-stoichiometric amount of the metal ions. Nonetheless, it is difficult to probe how those metal ions are distributed in the two identical subunits. By utilizing native mass spectrometry, we showed here that addition of a sub-stoichiometric copper/zinc ion to SOD1 led to the formation of a homodimer with a stochastic combination of the subunits binding 0, 1, and even 2 metal ions. We also found that the homodimer was able to bind four copper or four zinc ions, implying the binding of a copper and zinc ion at the canonical zinc and copper site, respectively. Such ambiguity in the metal quota and selectivity could be avoided when an intra-subunit disulfide bond in SOD1 was reduced before addition of the metal ions. Apo-SOD1 in the disulfide-reduced state was monomeric and was found to bind only one zinc ion per monomer. By binding a zinc ion, the disulfide-reduced SOD1 became conformationally compact and acquired the ability to dimerize. Based upon the results in vitro, we describe the pathway in vivo enabling SOD1 to bind copper and zinc ions with high accuracy in their quota and selectivity. A failure of correct metallation in SOD1 will also be discussed in relation to amyotrophic lateral sclerosis.

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A pathological link between dysregulated copper binding in Cu/Zn-superoxide dismutase and amyotrophic lateral sclerosis

Furukawa Y.

Journal of Clinical Biochemistry and Nutrition 71 ( 2 ) 73 - 77 2022

ISSN 09120009

Mutations in the gene coding Cu/Zn-superoxide dismutase (SOD1) are linked to a familial form of amyotrophic lateral sclerosis (ALS), and its pathological hallmark includes abnormal accumulation of mutant SOD1 proteins in spinal motorneurons. Mutant SOD1 proteins are considered to be susceptible to misfolding, resulting in the accumulation as oligomers/aggregates. While it remains obscure how and why SOD1 becomes misfolded under pathological conditions in vivo, the failure to bind a copper and zinc ion in SOD1 in vitro leads to the significant destabilization of its natively folded structure. Therefore, genetic and pharmacological attempts to promote the metal binding in mutant SOD1 could serve as an effective treatment of ALS. Here, I briefly review the copper and zinc binding process of SOD1 in vivo and discuss a copper chaperone for SOD1 as a potential target for developing ALS therapeutics.

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Stagnation of glymphatic interstitial fluid flow and delay in waste clearance in the SOD1-G93A mouse model of ALS.

Mikako Hirose, Mito Asano, Saori Watanabe-Matsumoto, Koji Yamanaka, Yoichiro Abe, Masato Yasui, Eiichi Tokuda, Yoshiaki Furukawa, Hidemi Misawa

Neuroscience research (Neuroscience Research) 171 74 - 82 2021.10

Accepted, ISSN 01680102

Overexpression and mislocalization of aquaporin-4 (AQP4) in the SOD1G93A mouse model of amyotrophic lateral sclerosis (ALS) have previously been reported. However, how alterations of AQP4 affect interstitial bulk flow in the brain and spinal cord, the so-called glymphatic system, is unclear. Here, we report an enhanced accumulation of disease-associated SOD1 species including SOD1 oligomers in SOD1G93A;AQP4-/- mice compared with SOD1G93A mice during ALS disease progression, as analyzed by sandwich ELISA. By directly injecting SOD1 oligomers into the spinal cord parenchyma, we observed a significantly larger delay in clearance of biotinylated or fluorescent-labeled SOD1 oligomers in AQP4-/- mice than in wild-type mice. Furthermore, when we injected the fluorescent-labeled tracer protein ovalbumin into the cisterna magna and analyzed the tracer distribution in the cervical spinal cord, approximately 35 % processing ability was found to be reduced in SOD1G93A mice compared to wild-type mice. These results suggest that the glymphatic system is abnormal and that waste clearance is delayed in SOD1G93A mice.

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A dual role of cysteine residues in the maturation of prokaryotic Cu/Zn-superoxide dismutase.

Yoshiaki Furukawa, Atsuko Shintani, Teppei Kokubo

Metallomics : integrated biometal science (Metallomics) 13 ( 9 ) 2021.09

Accepted, ISSN 17565901

Bacterial Cu/Zn-superoxide dismutase (SodC) is an enzyme catalyzing the disproportionation of superoxide radicals, to which the binding of copper and zinc ions and the formation of an intramolecular disulfide bond are essential. We previously showed that Escherichia coli SodC (SodC) was prone to spontaneous degradation in vivo in an immature form prior to the introduction of the disulfide bond. The post-translational maintenance involving the metal binding and the disulfide formation would thus control the stability as well as the enzymatic function of SodC; however, a mechanism of the SodC maturation remains obscure. Here, we show that the disulfide-reduced SodC can secure a copper ion as well as a zinc ion through the thiolate groups. Furthermore, the disulfide-reduced SodC was found to bind cuprous and cupric ions more tightly than SodC with the disulfide bond. The thiolate groups ligating the copper ion were then autooxidized to form the intramolecular disulfide bond, leading to the production of enzymatically active SodC. Based upon the experiments in vitro, therefore, we propose a mechanism for the activation of SodC, in which the conserved Cys residues play a dual role: the acquisition of a copper ion for the enzymatic activity and the formation of the disulfide bond for the structural stabilization.

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Good and bad of Cu/Zn-superoxide dismutase controlled by metal ions and disulfide bonds

Furukawa Y.

Chemistry Letters 50 ( 2 ) 331 - 341 2021

ISSN 03667022

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Does wild-type Cu/Zn-superoxide dismutase have pathogenic roles in amyotrophic lateral sclerosis?

Yoshiaki Furukawa, Eiichi Tokuda

Translational Neurodegeneration (Springer Science and Business Media LLC) 9 ( 1 ) 2020.08

Accepted

Amyotrophic lateral sclerosis (ALS) is characterized by adult-onset progressive degeneration of upper and lower motor neurons. Increasing numbers of genes are found to be associated with ALS; among those, the first identified gene, SOD1 coding a Cu/Zn-superoxide dismutase protein (SOD1), has been regarded as the gold standard in the research on a pathomechanism of ALS. Abnormal accumulation of misfolded SOD1 in affected spinal motor neurons has been established as a pathological hallmark of ALS caused by mutations in SOD1 (SOD1-ALS). Nonetheless, involvement of wild-type SOD1 remains quite controversial in the pathology of ALS with no SOD1 mutations (non-SOD1 ALS), which occupies more than 90% of total ALS cases. In vitro studies have revealed post-translationally controlled misfolding and aggregation of wild-type as well as of mutant SOD1 proteins; therefore, SOD1 proteins could be a therapeutic target not only in SOD1-ALS but also in more prevailing cases, non-SOD1 ALS. In order to search for evidence on misfolding and aggregation of wild-type SOD1 in vivo, we reviewed pathological studies using mouse models and patients and then summarized arguments for and against possible involvement of wild-type SOD1 in non-SOD1 ALS as well as in SOD1-ALS.

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Oxidative misfolding of Cu/Zn-superoxide dismutase triggered by non-canonical intramolecular disulfide formation.

Itsuki Anzai, Eiichi Tokuda, Sumika Handa, Hidemi Misawa, Shuji Akiyama, Yoshiaki Furukawa

Free radical biology & medicine (Free Radical Biology and Medicine) 147 187 - 199 2020.02

Accepted, ISSN 08915849

Misfolded Cu/Zn-superoxide dismutase (SOD1) is a pathological species in a subset of amyotrophic lateral sclerosis (ALS). Oxidative stress is known to increase in affected spinal cords of ALS and is thus considered to cause damages on SOD1 leading to the misfolding and aggregation. Despite this, it still remains elusive what triggers misfolding of SOD1 under oxidizing environment. Here, we show that a thiol group of Cys111 in SOD1 is oxidized to a sulfenic acid with hydrogen peroxide and reveal that further dissociation of the bound metal ions from the oxidized SOD1 allows another free Cys residue (Cys6) to nucleophilically attack the sulfenylated Cys111. As a result, an intra-molecular disulfide bond forms between Cys6 and Cys111. Such an abnormal SOD1 with the non-canonical disulfide bond was conformationally extended with significant cytotoxicity as well as high propensity to aggregate. Taken together, we propose a new model of SOD1 misfolding under oxidizing environment, in which formation of the non-canonical intramolecular disulfide bond plays a pivotal role.

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Wild-type Cu/Zn-superoxide dismutase is misfolded in cerebrospinal fluid of sporadic amyotrophic lateral sclerosis.

Eiichi Tokuda, Yo-Ichi Takei, Shinji Ohara, Noriko Fujiwara, Isao Hozumi, Yoshiaki Furukawa

Molecular neurodegeneration (Molecular Neurodegeneration) 14 ( 1 ) 42 - 42 2019.11

Accepted

BACKGROUND: A subset of familial forms of amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene coding Cu/Zn-superoxide dismutase (SOD1). Mutant SOD1 proteins are susceptible to misfolding and abnormally accumulated in spinal cord, which is most severely affected in ALS. It, however, remains quite controversial whether misfolding of wild-type SOD1 is involved in more prevalent sporadic ALS (sALS) cases without SOD1 mutations. METHODS: Cerebrospinal fluid (CSF) from patients including sALS as well as several other neurodegenerative diseases and non-neurodegenerative diseases was examined with an immunoprecipitation assay and a sandwich ELISA using antibodies specifically recognizing misfolded SOD1. RESULTS: We found that wild-type SOD1 was misfolded in CSF from all sALS cases examined in this study. The misfolded SOD1 was also detected in CSF from a subset of Parkinson's disease and progressive supranuclear palsy, albeit with smaller amounts than those in sALS. Furthermore, the CSF samples containing the misfolded SOD1 exhibited significant toxicity toward motor neuron-like NSC-34 cells, which was ameliorated by removal of the misfolded wild-type SOD1 with immunoprecipitation. CONCLUSIONS: Taken together, we propose that misfolding of wild-type SOD1 in CSF is a common pathological process of ALS cases regardless of SOD1 mutations.

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Sequence- and seed-structure-dependent polymorphic fibrils of alpha-synuclein.

Tanaka G, Yamanaka T, Furukawa Y, Kajimura N, Mitsuoka K, Nukina N

Biochimica et biophysica acta. Molecular basis of disease (Biochimica et Biophysica Acta - Molecular Basis of Disease) 1865 ( 6 ) 1410 - 1420 2019.06

Accepted, ISSN 09254439

Synucleinopathies comprise a diverse group of neurodegenerative diseases including Parkinson's disease (PD), dementia with Lewy bodies, and multiple system atrophy. These share a common pathological feature, the deposition of alpha-synuclein (a-syn) in neurons or oligodendroglia. A-syn is highly conserved in vertebrates, but the primary sequence of mouse a-syn differs from that of human at seven positions. However, structural differences of their aggregates remain to be fully characterized. In this study, we found that human and mouse a-syn aggregated in vitro formed morphologically distinct amyloid fibrils exhibiting twisted and straight structures, respectively. Furthermore, we identified different protease-resistant core regions, long and short, in human and mouse a-syn aggregates. Interestingly, among the seven unconserved amino acids, only A53T substitution, one of the familial PD mutations, was responsible for structural conversion to the straight-type. Finally, we checked whether the structural differences are transmissible by seeding and found that human a-syn seeded with A53T aggregates formed straight-type fibrils with short protease-resistant cores. These results suggest that a-syn aggregates form sequence-dependent polymorphic fibrils upon spontaneous aggregation but become seed structure-dependent upon seeding.

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Biochemical and morphological classification of disease-associated alpha-synuclein mutants aggregates.

Tanaka G, Yamanaka T, Furukawa Y, Kajimura N, Mitsuoka K, Nukina N

Biochemical and biophysical research communications (Biochemical and Biophysical Research Communications) 508 ( 3 ) 729 - 734 2019.01

Accepted, ISSN 0006291X

Alpha-synuclein (a-syn) aggregation in brain is implicated in several synucleinopathies, including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Until date, at least six disease-associated mutations in a-syn (namely A30P, E46K, H50Q, G51D, A53T, and A53E) are known to cause dominantly inherited familial forms of synucleinopathies. Previous studies using recombinant proteins have reported that a subset of disease-associated mutants show higher aggregation propensities and form spectroscopically distinguishable aggregates compared to wild-type (WT). However, morphological and biochemical comparison of the aggregates for all disease-associated a-syn mutants have not yet been performed. In this study, we performed electron microscopic examination, guanidinium hydrochloride (GdnHCl) denaturation, and protease digestion to classify the aggregates from their respective point mutations. Using electron microscopy we observed variations of amyloid fibrillar morphologies among the aggregates of a-syn mutants, mainly categorized into two groups: twisted fibrils observed for both WT and E46K while straight fibrils for the other mutants. GdnHCl denaturation experiments revealed the a-syn mutants except for E46K were more resistant than WT against the denaturation. Mass spectrometry analysis of protease-treated aggregates showed a variety of protease-resistant cores, which may correspond to their morphological properties. The difference of their properties could be implicated in the clinicopathological difference of synucleinopathies with those mutations.

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Prion-like properties of misfolded Cu/Zn-superoxide dismutase in amyotrophic lateral sclerosis: Update and perspectives

Tokuda E., Marklund S., Furukawa Y.

Yakugaku Zasshi (Yakugaku Zasshi) 139 ( 7 ) 1015 - 1019 2019

ISSN 00316903

© 2019 The Pharmaceutical Society of Japan. Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease that is characterized by the loss of motor neurons, which results in progressive muscle atrophy. The pathology spreads from the initial site of onset to contiguous anatomic regions. Mutations in the gene encoding Cu/Zn-superoxide dismutase (SOD1) have been identified in a dominantly inherited form of ALS (ALS-SOD1). A major hallmark of ALS-SOD1 is the abnormal accumulation of conformationally aberrant SOD1 protein (i.e., misfolded SOD1) within motor neurons. Emerging experimental evidence has suggested that misfolded proteins associated with neurodegenerative diseases exhibit prion-like properties, i.e., misfolded proteins act as conformational templates that convert normal proteins into a pathogenic form. Possibly as a result of this prion-like self-propagation property, misfolded forms of pathological proteins are considered to accumulate in the central nervous system and cause neurodegeneration. In this article, we review recent evidence for the role of prion-like mechanisms in ALS-SOD1. In particular, we discuss the propensity of misfolded SOD1 to act as a pathological seed, spread between cells, and propagate neuroanatomically.

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Elimination of TDP-43 inclusions linked to amyotrophic lateral sclerosis by a misfolding-specific intrabody with dual proteolytic signals

Yoshitaka Tamaki, Akemi Shodai, Toshifumi Morimura, Ryota Hikiami, Sumio Minamiyama, Takashi Ayaki, Ikuo Tooyama, Yoshiaki Furukawa, Ryosuke Takahashi, Makoto Urushitani

Scientific Reports (Scientific Reports) 8 ( 1 ) 6030 2018.12

Accepted, ISSN 2045-2322

© 2018 The Author(s). Aggregation of TAR DNA-binding protein of 43 kDa (TDP-43) is implicated in the pathogenesis of sporadic and certain familial forms of amyotrophic lateral sclerosis (ALS), suggesting elimination of TDP-43 aggregates as a possible therapeutic strategy. Here we generated and investigated a single-chain variable fragment (scFv) derived from the 3B12A monoclonal antibody (MAb) that recognises D247 of the TDP-43 nuclear export signal, an epitope masked in the physiological state. In transfected HEK293A cells, 3B12A scFv recapitulated the affinity of the full-length MAb to mislocalised TDP-43 with a defective nuclear localising signal and to a TDP-43 inclusion mimic with cysteine-to-serine substitution at RRM1. Moreover, 3B12A scFv accelerated proteasome-mediated degradation of aggregated TDP-43, likely due to an endogenous PEST-like proteolytic signal sequence in the VH domain CDR2 region. Addition of the chaperone-mediated autophagy (CMA)-related signal to 3B12A scFv induced HSP70 transcription, further enhancing TDP-43 aggregate clearance and cell viability. The 3B12A scFv also reduced TDP-43 aggregates in embryonic mouse brain following in utero electroporation while causing no overt postnatal brain pathology or developmental anomalies. These results suggest that a misfolding-specific intrabody prone to synergistic proteolysis by proteasomal and autophagic pathways is a promising strategy for mitigation of TDP-43 proteinopathy in ALS.

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Cu/Zn-superoxide dismutase forms fibrillar hydrogels in a pH-dependent manner via a water-rich extended intermediate state.

Fujiwara N, Wagatsuma M, Oba N, Yoshihara D, Tokuda E, Sakiyama H, Eguchi H, Ichihashi M, Furukawa Y, Inoue T, Suzuki K

PloS one (Public Library of Science (PLoS)) 13 ( 10 ) e0205090 - e0205090 2018.10

Accepted

© 2018 Fujiwara et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Under certain conditions, amyloid-like fibrils can develop into three-dimensional networks and form hydrogels by a self-assembly process. When Cu/Zn superoxide dismutase (SOD1), an anti-oxidative enzyme, undergoes misfolding, fibrillar aggregates are formed, which are a hallmark of a certain form of familial amyotrophic lateral sclerosis (ALS). However, the issue of whether SOD1 fibrils can be assembled into hydrogels remains to be tested. Here, we show that the SOD1 polypeptides undergo hydrogelation accompanied by the formation of thioflavin T-positive fibrils at pH 3.0 and 4.0, but not at pH 5.0 where precipitates are formed. The results of viscoelastic analyses indicate that the properties of SOD1 hydrogels (2%) were similar to and slightly more fragile than a 0.25% agarose gel. In addition, monitoring by a quartz crystal microbalance with admittance analysis showed that the denaturing of immobilized SOD1 on a sensor under the hydrogelation conditions at pH 3.0 and 4.0 resulted in an increase in the effective acoustic thickness from ~3.3 nm (a folded rigid form) to ~50 and ~100 nm (an extended water-rich state), respectively. In contrast, when SOD1 was denatured under the same conditions at pH 5.0, a compact water-poor state with an effective acoustic thickness of ~10 nm was formed. The addition of physiological concentrations of NaCl to the pH 4.0 sample induced a further extension of the SOD1 with larger amounts of water molecules (with an effective acoustic thickness of ~200 nm) but suppressed hydrogel formation. These results suggest that different denatured intermediate states of the protein before self-assembly play a major role in determining the characteristics of the resulting aggregates and that a conformational change to a suitable level of extended water-rich intermediate state before and/or during intermolecular assembling is required for fibrillation and hydrogelation in the case of globular proteins.

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Rapid dissemination of alpha-synuclein seeds through neural circuits in an in-vivo prion-like seeding experiment.

Ayami Okuzumi, Masaru Kurosawa, Taku Hatano, Masashi Takanashi, Shuuko Nojiri, Takeshi Fukuhara, Tomoyuki Yamanaka, Haruko Miyazaki, Saki Yoshinaga, Yoshiaki Furukawa, Tomomi Shimogori, Nobutaka Hattori, Nobuyuki Nukina

Acta neuropathologica communications (Acta neuropathologica communications) 6 ( 1 ) 96 - 96 2018.09

Accepted

Accumulating evidence suggests that the lesions of Parkinson's disease (PD) expand due to transneuronal spreading of fibrils composed of misfolded alpha-synuclein (a-syn), over the course of 5-10 years. However, the precise mechanisms and the processes underlying the spread of these fibril seeds have not been clarified in vivo. Here, we investigated the speed of a-syn transmission, which has not been a focus of previous a-syn transmission experiments, and whether a-syn pathologies spread in a neural circuit-dependent manner in the mouse brain. We injected a-syn preformed fibrils (PFFs), which are seeds for the propagation of a-syn deposits, either before or after callosotomy, to disconnect bilateral hemispheric connections. In mice that underwent callosotomy before the injection, the propagation of a-syn pathology to the contralateral hemisphere was clearly reduced. In contrast, mice that underwent callosotomy 24 h after a-syn PFFs injection showed a-syn pathology similar to that seen in mice without callosotomy. These results suggest that a-syn seeds are rapidly disseminated through neuronal circuits immediately after seed injection, in a prion-like seeding experiment in vivo, although it is believed that clinical a-syn pathologies take years to spread throughout the brain. In addition, we found that botulinum toxin B blocked the transsynaptic transmission of a-syn seeds by specifically inactivating the synaptic vesicle fusion machinery. This study offers a novel concept regarding a-syn propagation, based on the Braak hypothesis, and also cautions that experimental transmission systems may be examining a unique type of transmission, which differs from the clinical disease state.

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Identification of a novel zinc-binding protein, C1orf123, as an interactor with a heavy metal-associated domain.

Furukawa Y, Lim C, Tosha T, Yoshida K, Hagai T, Akiyama S, Watanabe S, Nakagome K, Shiro Y

PloS one (PLoS ONE) 13 ( 9 ) e0204355 2018.09

Accepted

© 2018 Furukawa et al.This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Heavy metal-associated (HMA) domains bind metal ions at its Cys-x-x-Cys (CxxC) motif and constitute an intracellular network for trafficking of metal ions for utilization and detoxification. We thus expect that novel metalloproteins can be identified by screening proteins interacting with a HMA domain. In this study, we performed yeast two-hybrid screening of the human proteome and found an uncharacterized protein encoded as open reading frame 123 in chromosome 1 (C1orf123) that can interact specifically with the HMA domain of a copper chaperone for superoxide dismutase (CCSdI). Our X-ray structural analysis of C1orf123 further revealed that it binds a Zn 2+ ion in a tetrahedral coordination with four thiolate groups from two conserved CxxC motifs. For the interaction between C1orf123 and CCSdI, the CxxC motifs in both C1orf123 and CCSdI were required, implying metal-mediated interaction through the CxxC motifs. Notably, C1orf123 did not interact with several other HMA domains containing CxxC motifs, supporting high specificity in the interaction between C1orf123 and CCSdI. Based upon these results, we further discuss functional and structural significance of the interaction between C1orf123 and CCS.

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A copper-deficient form of mutant Cu/Zn-superoxide dismutase as an early pathological species in amyotrophic lateral sclerosis

Eiichi Tokuda, Takao Nomura, Shinji Ohara, Seiji Watanabe, Koji Yamanaka, Yuta Morisaki, Hidemi Misawa, Yoshiaki Furukawa

Biochimica et Biophysica Acta - Molecular Basis of Disease (Biochimica et Biophysica Acta - Molecular Basis of Disease) 1864 ( 6 ) 2119 - 2130 2018.06

Accepted, ISSN 09254439

© 2018 Elsevier B.V. Dominant mutations in the gene encoding copper and zinc-binding superoxide dismutase (SOD1) cause amyotrophic lateral sclerosis (ALS). Abnormal accumulation of misfolded SOD1 proteins in spinal motoneurons is a major pathological hallmark in SOD1-related ALS. Dissociation of copper and/or zinc ions from SOD1 has been shown to trigger the protein aggregation/oligomerization in vitro, but the pathological contribution of such metal dissociation to the SOD1 misfolding still remains obscure. Here, we tested the relevance of the metal-deficient SOD1 in the misfolding in vivo by developing a novel antibody (anti-apoSOD), which exclusively recognized mutant SOD1 deficient in metal ions at its copper-binding site. Notably, anti-apoSOD-reactive species were detected specifically in the spinal cords of the ALS model mice only at their early pre-symptomatic stages but not at the end stage of the disease. The cerebrospinal fluid as well as the spinal cord homogenate of one SOD1-ALS patient also contained the anti-apoSOD-reactive species. Our results thus suggest that metal-deficiency in mutant SOD1 at its copper-binding site is one of the earliest pathological features in SOD1-ALS.

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Pathological roles of copper dynamics in amyotrophic lateral sclerosis

古川良明

生化学 (日本生化学会) 90 ( 3 ) 311 - 319 2018

ISSN 00371017

An essential role of N-terminal domain of copper chaperone in the enzymatic activation of Cu/Zn-superoxide dismutase

Mami Fukuoka, Eiichi Tokuda, Kenta Nakagome, Zhiliang Wu, Isao Nagano, Yoshiaki Furukawa

JOURNAL OF INORGANIC BIOCHEMISTRY 175 208 - 216 2017.10

Accepted, ISSN 0162-0134

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Regulatory Role of RNA Chaperone TDP-43 for RNA Misfolding and Repeat-Associated Translation in SCA31

Taro Ishiguro, Nozomu Sato, Morio Ueyama, Nobuhiro Fujikake, Chantal Sellier, Akemi Kanegami, Eiichi Tokuda, Bita Zamiri, Terence Gall-Duncan, Mila Mirceta, Yoshiaki Furukawa, Takanori Yokota, Keiji Wada, J. Paul Taylor, Christopher E. Pearson, Nicolas Charlet-Berguerand, Hidehiro Mizusawa, Yoshitaka Nagai, Kinya Ishikawa

NEURON 94 ( 1 ) 108 - + 2017.04

Accepted, ISSN 0896-6273

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A misfolded dimer of Cu/Zn-superoxide dismutase leading to pathological oligomerization in amyotrophic lateral sclerosis

Itsuki Anzai, Eiichi Tokuda, Atsushi Mukaiyama, Shuji Akiyama, Fumito Endo, Koji Yamanaka, Hidemi Misawa, Yoshiaki Furukawa

PROTEIN SCIENCE 26 ( 3 ) 484 - 496 2017.03

Accepted, ISSN 0961-8368

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Immunochemical characterization on pathological oligomers of mutant Cu/Znsuperoxide dismutase in amyotrophic lateral sclerosis

Tokuda, Eiichi, Anzai, Itsuki, Nomura, Takao, Toichi, Keisuke, Watanabe, Masahiko, Ohara, Shinji, Watanabe, Seiji, Yamanaka, Koji, Morisaki, Yuta, Misawa, Hidemi, Furukawa, Yoshiaki

MOLECULAR NEURODEGENERATION 12 ( 1 ) 2 2017.01

Research paper (scientific journal), Accepted, ISSN 1750-1326

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Aggregation of FET proteins as a pathological change in amyotrophic lateral sclerosis

Yoshiaki Furukawa, Eiichi Tokuda

Advances in Experimental Medicine and Biology (Springer New York LLC) 925 1 - 12 2017

Accepted, ISSN 2214-8019

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Screening of drugs inhibiting in vitro oligomerization of Cu/Zn-superoxide dismutase with a mutation causing amyotrophic lateral sclerosis

Itsuki Anzai, Keisuke Toichi, Eiichi Tokuda, Atsushi Mukaiyama, Shuji Akiyama, Yoshiaki Furukawa

Frontiers in Molecular Biosciences (Frontiers Media S.A.) 3 ( AUG ) 40 2016.08

Accepted, ISSN 2296-889X

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Copper Homeostasis as a Therapeutic Target in Amyotrophic Lateral Sclerosis with SOD1 Mutations

Tokuda, Eiichi, Furukawa, Yoshiaki

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 17 ( 5 ) 847 - 867 2016.05

Research paper (scientific journal), Accepted, ISSN 1422-0067

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Conformational Disorder of the Most Immature Cu, Zn-Superoxide Dismutase Leading to Amyotrophic Lateral Sclerosis

Furukawa, Yoshiaki, Anzai, Itsuki, Akiyama, Shuji, Imai, Mizue, Cruz, Fatima Joy C., Saio, Tomohide, Nagasawa, Kenichi, Nomura, Takao, Ishimori, Koichiro

JOURNAL OF BIOLOGICAL CHEMISTRY 291 ( 8 ) 4144 - 4155 2016.02

Research paper (scientific journal), ISSN 0021-9258

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A molecular mechanism realizing sequence-specific recognition of nucleic acids by TDP-43

Furukawa, Yoshiaki, Suzuki, Yoh, Fukuoka, Mami, Nagasawa, Kenichi, Nakagome, Kenta, Shimizu, Hideaki, Mukaiyama, Atsushi, Akiyama, Shuji

SCIENTIFIC REPORTS 6 4144 - 4155 2016.02

Research paper (scientific journal), ISSN 2045-2322

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Close Up実験法 Series258 タンパク質を再現性よく凝集させるコツ

古川良明

実験医学 ((株)羊土社) 33 ( 1 ) 93 - 97 2015.01

ISSN 0288-5514

神経・非神経細胞ネットワークと脳内環境 13.筋萎縮性側索硬化症におけるタンパク質の線維化とシーディング現象

古川良明

遺伝子医学MOOK ( 26 ) 153 - 157 2014.11

ISSN 1349-2527

A Primary Role for Disulfide Formation in the Productive Folding of Prokaryotic Cu,Zn-superoxide Dismutase

Yasuyuki Sakurai, Itsuki Anzai, Yoshiaki Furukawa

JOURNAL OF BIOLOGICAL CHEMISTRY 289 ( 29 ) 20139 - 20149 2014.07

Accepted, ISSN 0021-9258

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A seeded propagation of Cu, Zn-superoxide dismutase aggregates in amyotrophic lateral sclerosis

Mariko Ogawa, Yoshiaki Furukawa

FRONTIERS IN CELLULAR NEUROSCIENCE 8 83 2014.03

Accepted, ISSN 1662-5102

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Intranuclear Aggregation of Mutant FUS/TLS as a Molecular Pathomechanism of Amyotrophic Lateral Sclerosis

Takao Nomura, Shoji Watanabe, Kumi Kaneko, Koji Yamanaka, Nobuyuki Nukina, Yoshiaki Furukawa

JOURNAL OF BIOLOGICAL CHEMISTRY 289 ( 2 ) 1192 - 1202 2014.01

Accepted, ISSN 0021-9258

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Redox environment is an intracellular factor to operate distinct pathways for aggregation of Cu,Zn-superoxide dismutase in amyotrophic lateral sclerosis

Yoshiaki Furukawa

FRONTIERS IN CELLULAR NEUROSCIENCE 7 240 2013.11

Accepted, ISSN 1662-5102

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Functional diversity of protein fibrillar aggregates from physiology to RNA granules to neurodegenerative diseases

Yoshiaki Furukawa, Nobuyuki Nukina

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE 1832 ( 8 ) 1271 - 1278 2013.08

Accepted, ISSN 0925-4439

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Intracellular seeded aggregation of mutant Cu,Zn-superoxide dismutase associated with amyotrophic lateral sclerosis

Yoshiaki Furukawa, Kumi Kaneko, Shoji Watanabe, Koji Yamanaka, Nobuyuki Nukina

FEBS LETTERS 587 ( 16 ) 2500 - 2505 2013.08

Accepted, ISSN 0014-5793

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Disulfide scrambling describes the oligomer formation of superoxide dismutase (SOD1) proteins in the familial form of amyotrophic lateral sclerosis

Keisuke Toichi, Koji Yamanaka, Yoshiaki Furukawa

Journal of Biological Chemistry 288 ( 7 ) 4970 - 4980 2013.02

Accepted, ISSN 0021-9258

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Post-aggregation Oxidation of Mutant Huntingtin Controls the Interactions between Aggregates

Yasushi Mitomi, Takao Nomura, Masaru Kurosawa, Nobuyuki Nukina, Yoshiaki Furukawa

JOURNAL OF BIOLOGICAL CHEMISTRY 287 ( 41 ) 34764 - 34775 2012.10

Accepted, ISSN 0021-9258

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Local Unfolding of Cu, Zn Superoxide Dismutase Monomer Determines the Morphology of Fibrillar Aggregates

Feng Ding, Yoshiaki Furukawa, Nobuyuki Nukina, Nikolay V. Dokholyan

JOURNAL OF MOLECULAR BIOLOGY 421 ( 4-5 ) 548 - 560 2012.08

Accepted, ISSN 0022-2836

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Pathological roles of wild-type Cu, Zn-superoxide dismutase in amyotrophic lateral sclerosis

Yoshiaki Furukawa

Neurology Research International 2012 323261 2012

Accepted, ISSN 2090-1852

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Molecular properties of TAR DNA binding protein-43 fragments are dependent upon its cleavage site

Yoshiaki Furukawa, Kumi Kaneko, Nobuyuki Nukina

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE 1812 ( 12 ) 1577 - 1583 2011.12

Accepted, ISSN 0925-4439

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Tau Protein Assembles into Isoform- and Disulfide-dependent Polymorphic Fibrils with Distinct Structural Properties

Yoshiaki Furukawa, Kumi Kaneko, Nobuyuki Nukina

JOURNAL OF BIOLOGICAL CHEMISTRY 286 ( 31 ) 27236 - 27246 2011.08

Accepted, ISSN 0021-9258

A Seeding Reaction Recapitulates Intracellular Formation of Sarkosyl-insoluble Transactivation Response Element (TAR) DNA-binding Protein-43 Inclusions

Yoshiaki Furukawa, Kumi Kaneko, Shoji Watanabe, Koji Yamanaka, Nobuyuki Nukina

JOURNAL OF BIOLOGICAL CHEMISTRY 286 ( 21 ) 18664 - 18672 2011.05

Accepted, ISSN 0021-9258

Mutation-dependent Polymorphism of Cu,Zn-Superoxide Dismutase Aggregates in the Familial Form of Amyotrophic Lateral Sclerosis

Yoshiaki Furukawa, Kumi Kaneko, Koji Yamanaka, Nobuyuki Nukina

JOURNAL OF BIOLOGICAL CHEMISTRY 285 ( 29 ) 22221 - 22231 2010.07

Accepted, ISSN 0021-9258

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Mutant huntingtin fragment selectively suppresses Brn-2 POU domain transcription factor to mediate hypothalamic cell dysfunction

Tomoyuki Yamanaka, Asako Tosaki, Haruko Miyazaki, Masaru Kurosawa, Yoshiaki Furukawa, Mizuki Yamada, Nobuyuki Nukina

HUMAN MOLECULAR GENETICS 19 ( 11 ) 2099 - 2112 2010.06

Accepted, ISSN 0964-6906

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Cross-Seeding Fibrillation of Q/N-Rich Proteins Offers New Pathomechanism of Polyglutamine Diseases

Yoshiaki Furukawa, Kumi Kaneko, Gen Matsumoto, Masaru Kurosawa, Nobuyuki Nukina

JOURNAL OF NEUROSCIENCE 29 ( 16 ) 5153 - 5162 2009.04

Accepted, ISSN 0270-6474

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Complete loss of post-translational modifications triggers fibrillar aggregation of SOD1 in the familial form of amyotrophic lateral sclerosis

Yoshiaki Furukawa, Kumi Kaneko, Koji Yamanaka, Thomas V. O'Halloran, Nobuyuki Nukina

JOURNAL OF BIOLOGICAL CHEMISTRY 283 ( 35 ) 24167 - 24176 2008.08

Accepted, ISSN 0021-9258

RNA-binding protein TLS is a major nuclear aggregate-interacting protein in huntingtin exon 1 with expanded polyglutamine-expressing cells

Hiroshi Doi, Kazumasa Okamura, Peter O. Bauer, Yoshiaki Furukawa, Hideaki Shimizu, Masaru Kurosawa, Yoko Machida, Haruko Miyazaki, Kenichi Mitsui, Yoshiyuki Kuroiwa, Nobuyuki Nukina

JOURNAL OF BIOLOGICAL CHEMISTRY 283 ( 10 ) 6489 - 6500 2008.03

Accepted, ISSN 0021-9258

Conversion to the amyotrophic lateral sclerosis phenotype is associated with intermolecular linked insoluble aggregates of SOD1 in mitochondria

HX Deng, Y Shi, Y Furukawa, H Zhai, RG Fu, ED Liu, GH Gorrie, MS Khan, WY Hung, EH Bigio, T Lukas, MC Dal Canto, TV O'Halloran, T Siddique

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 103 ( 18 ) 7142 - 7147 2006.05

Accepted, ISSN 0027-8424

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Posttranslational modifications in Cu,Zn-superoxide dismutase and mutations associated with arnyotrophic lateral sclerosis

Yoshiaki Furukawa, Thomas V. O'Halloran

ANTIOXIDANTS & REDOX SIGNALING 8 ( 5-6 ) 847 - 867 2006.05

Accepted, ISSN 1523-0864

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Disulfide cross-linked protein represents a significant fraction of ALS-associated Cu, Zn-superoxide dismutase aggregates in spinal cords of model mice

Y Furukawa, RG Fu, HX Deng, T Siddique, TV O'Halloran

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 103 ( 18 ) 7148 - 7153 2006.05

Accepted, ISSN 0027-8424

Amyotrophic lateral sclerosis mutations have the greatest destabilizing effect on the Apo- and reduced form of SOD1, leading to unfolding and oxidative aggregation

Y Furukawa, TV O'Halloran

JOURNAL OF BIOLOGICAL CHEMISTRY 280 ( 17 ) 17266 - 17274 2005.04

Accepted, ISSN 0021-9258

Electron transfer reaction in a single protein molecule observed by total internal reflection fluorescence microscopy

Y Furukawa, T Ban, D Hamada, K Ishimori, Y Goto, Morishima, I

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 127 ( 7 ) 2098 - 2103 2005.02

Accepted, ISSN 0002-7863

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The unusually stable quaternary structure of human Cu,Zn-superoxide dismutase 1 is controlled by both metal occupancy and disulfide status

F Arnesano, L Banci, Bertini, I, M Martinelli, Y Furukawa, TV O'Halloran

JOURNAL OF BIOLOGICAL CHEMISTRY 279 ( 46 ) 47998 - 48003 2004.11

Accepted, ISSN 0021-9258

Oxygen-induced maturation of SOD1: a key role for disulfide formation by the copper chaperone CCS

Y Furukawa, AS Torres, TV O'Halloran

EMBO JOURNAL 23 ( 14 ) 2872 - 2881 2004.07

Accepted, ISSN 0261-4189

Factors controlling the uptake of yeast copper/zinc superoxide dismutase into mitochondria

LS Field, Y Furukawa, TV O'Halloran, VC Culotta

JOURNAL OF BIOLOGICAL CHEMISTRY 278 ( 30 ) 28052 - 28059 2003.07

Accepted, ISSN 0021-9258

Oxidation-state-dependent protein docking between cytochrome c and cytochrome b(5): High-pressure laser flash photolysis study

Y Furukawa, K Ishimori, Morishima, I

BIOCHEMISTRY 41 ( 31 ) 9824 - 9832 2002.08

Accepted, ISSN 0006-2960

Investigation of the electron-transfer mechanism by cross-linking between Zn-substituted myoglobin and cytochrome b(5)

Y Furukawa, F Matsuda, K Ishimori, Morishima, I

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 124 ( 15 ) 4008 - 4019 2002.04

Accepted, ISSN 0002-7863

Pressure Effects on the Intramolecular Electron Transfer Reactions in Hemoproteins

Yoshiaki Furukawa, Yoichi Sugiyama, Satoshi Takahashi, Koichiro Ishimori, Isao Morishima

(Springer Nature) 187 2002

Accepted

Access to Document (DOI)

The role of water molecules in the association of cytochrome P450cam with putidaredoxin - An osmotic pressure study

Y Furukawa, Morishima, I

JOURNAL OF BIOLOGICAL CHEMISTRY 276 ( 16 ) 12983 - 12990 2001.04

Accepted, ISSN 0021-9258

Electron transfer reactions in Zn-substituted cytochrome P450cam

Y Furukawa, K Ishimori, Morishima, I

BIOCHEMISTRY 39 ( 36 ) 10996 - 11004 2000.09

Accepted, ISSN 0006-2960

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Pressure dependence of the intramolecular electron transfer reaction in myoglobin reinvestigated

Y Furukawa, K Ishimori, Morishima, I

JOURNAL OF PHYSICAL CHEMISTRY B 104 ( 8 ) 1817 - 1825 2000.03

ISSN 1089-5647

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