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

Noriko Fujiwara, Michiru Wagatsuma, Naoto Oba, Daisaku Yoshihara, Eiichi Tokuda, Haruhiko Sakiyama, Hironobu Eguchi, Motoko Ichihashi, Yoshiaki Furukawa, Tadashi Inoue, Keiichiro Suzuki

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Article numbere0205090
JournalPLoS One
Volume13
Issue number10
DOIs
Publication statusPublished - 2018 Oct 1

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Hydrogels
hydrocolloids
superoxide dismutase
Acoustics
Water
acoustics
Self assembly
protein aggregates
water
Quartz Crystal Microbalance Techniques
Hydrogel
Quartz crystal microbalances
Amyloid
Sepharose
Precipitates
quartz
amyloid
Proteins
Gels
agarose

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Fujiwara, N., Wagatsuma, M., Oba, N., Yoshihara, D., Tokuda, E., Sakiyama, H., ... Suzuki, K. (2018). Cu/Zn-superoxide dismutase forms fibrillar hydrogels in a pH-dependent manner via a water-rich extended intermediate state. PLoS One, 13(10), [e0205090]. https://doi.org/10.1371/journal.pone.0205090

Cu/Zn-superoxide dismutase forms fibrillar hydrogels in a pH-dependent manner via a water-rich extended intermediate state. / Fujiwara, Noriko; Wagatsuma, Michiru; Oba, Naoto; Yoshihara, Daisaku; Tokuda, Eiichi; Sakiyama, Haruhiko; Eguchi, Hironobu; Ichihashi, Motoko; Furukawa, Yoshiaki; Inoue, Tadashi; Suzuki, Keiichiro.

In: PLoS One, Vol. 13, No. 10, e0205090, 01.10.2018.

Research output: Contribution to journalArticle

Fujiwara, N, Wagatsuma, M, Oba, N, Yoshihara, D, Tokuda, E, Sakiyama, H, Eguchi, H, Ichihashi, M, Furukawa, Y, Inoue, T & Suzuki, K 2018, 'Cu/Zn-superoxide dismutase forms fibrillar hydrogels in a pH-dependent manner via a water-rich extended intermediate state', PLoS One, vol. 13, no. 10, e0205090. https://doi.org/10.1371/journal.pone.0205090
Fujiwara, Noriko ; Wagatsuma, Michiru ; Oba, Naoto ; Yoshihara, Daisaku ; Tokuda, Eiichi ; Sakiyama, Haruhiko ; Eguchi, Hironobu ; Ichihashi, Motoko ; Furukawa, Yoshiaki ; Inoue, Tadashi ; Suzuki, Keiichiro. / Cu/Zn-superoxide dismutase forms fibrillar hydrogels in a pH-dependent manner via a water-rich extended intermediate state. In: PLoS One. 2018 ; Vol. 13, No. 10.
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