Oligomerization of Cu,Zn-superoxide dismutase (SOD1) by Docosahexaenoic acid and its hydroperoxides in vitro: Aggregation dependence on fatty acid unsaturation and thiols

Patricia Postilione Appolinário, Danilo Bilches Medinas, Adriano B. Chaves-Filho, Thiago C. Genaro-Mattos, José Renato Rosa Cussiol, Luis Eduardo Soares Netto, Ohara Augusto, Sayuri Miyamoto

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Docosahexaenoic acid (C22:6, n-3, DHA) is a polyunsaturated fatty acid highly enriched in the brain. This fatty acid can be easily oxidized yielding hydroperoxides as primary products. Cu, Zn-Superoxide dismutase (SOD1) aggregation is a common hallmark of Amyotrophic Lateral Sclerosis (ALS) and the molecular mechanisms behind their formation are not completely understood. Here we investigated the effect of DHA and its hydroperoxides (DHAOOH) on human SOD1 oligomerization in vitro. DHA induced the formation of highmolecularweight (HMW) SOD1 species (>700 kDa). Aggregation was dependent on free thiols and occurred primarily with the protein in its apo-form. SOD1 incubation with DHA was accompanied by changes in protein structure leading to exposure of protein hydrophobic patches and formation of non-amyloid aggregates. Site-directed mutagenesis studies demonstrated that Cys 6 and Cys 111 in wild-type and Cys 6 in ALS-linked G93A mutant are required for aggregation. In contrast, DHAOOH did not induce HMW species formation but promoted abnormal covalent dimerization of apo-SOD1 that was resistant to SDS and thiol reductants. Overall, our data demonstrate that DHA and DHAOOH induce distinct types of apo-SOD1 oligomerization leading to the formation of HMW and low-molecularweight species, respectively.

Original languageEnglish (US)
Article numbere0125146
JournalPloS one
Volume10
Issue number4
DOIs
StatePublished - Apr 1 2015

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Oligomerization
hydroperoxides
thiols
docosahexaenoic acid
Sulfhydryl Compounds
superoxide dismutase
Fatty Acids
Agglomeration
fatty acids
Hydrogen Peroxide
reducing agents
dimerization
site-directed mutagenesis
protein aggregates
protein structure
polyunsaturated fatty acids
Mutagenesis
Proteins
Dimerization
proteins

ASJC Scopus subject areas

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

Cite this

Oligomerization of Cu,Zn-superoxide dismutase (SOD1) by Docosahexaenoic acid and its hydroperoxides in vitro : Aggregation dependence on fatty acid unsaturation and thiols. / Appolinário, Patricia Postilione; Medinas, Danilo Bilches; Chaves-Filho, Adriano B.; Genaro-Mattos, Thiago C.; Cussiol, José Renato Rosa; Netto, Luis Eduardo Soares; Augusto, Ohara; Miyamoto, Sayuri.

In: PloS one, Vol. 10, No. 4, e0125146, 01.04.2015.

Research output: Contribution to journalArticle

Appolinário, Patricia Postilione ; Medinas, Danilo Bilches ; Chaves-Filho, Adriano B. ; Genaro-Mattos, Thiago C. ; Cussiol, José Renato Rosa ; Netto, Luis Eduardo Soares ; Augusto, Ohara ; Miyamoto, Sayuri. / Oligomerization of Cu,Zn-superoxide dismutase (SOD1) by Docosahexaenoic acid and its hydroperoxides in vitro : Aggregation dependence on fatty acid unsaturation and thiols. In: PloS one. 2015 ; Vol. 10, No. 4.
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