Amino acids of importance for the antioxidant activity of human serum albumin as revealed by recombinant mutants and genetic variants

Makoto Anraku, Rina Shintomo, Kazuaki Taguchi, Ulrich Kragh-Hansen, Toshiya Kai, Toru Maruyama, Masaki Otagiri

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Aims: To determine molecular information about the antioxidant properties of human serum albumin, which is an important extracellular antioxidant. To obtain this information, we studied this function of the protein by using H2O2 as the representative reactive oxygen species and two recombinant mutants and ten genetic variants with single-residue mutations. Main methods: The antioxidant capabilities of the isoforms were registered as their ability to diminish the H2O2-induced conversion of dihydrorhodamine 123 to rhodamine 123, which can emit fluorescence at 536 nm. Structural properties were examined by circular dichroism and SDS-PAGE. Key findings: Cysteine residues are important for the antioxidant function, but their effect depends on their position in the protein, with Cys410 > Cys34 ∼ Cys169 (when not involved in forming a disulfide bond). Likewise, the substitution of a glutamic acid at position 122 or 541, but not at 240 or 560, improves the antioxidant effect, perhaps by making the methionine residues in their vicinity, Met123 and Met548, respectively, more accessible for the oxidant. A lysine at position 505, but not at 82 or 570, decreases the oxidative effect. Finally, the mutations D269G and K276N had no effect. In certain cases, albumin acts as a sacrificial antioxidant, as in the case of the mutants C34S and, in particular, R410C and E505K. Significance: The information gained is of protein chemical relevance, but it may also be helpful in understanding the function of proteins that act as antioxidants in biological systems subjected to oxidative stress in conditions such as inflammation and aging.

Original languageEnglish
Pages (from-to)36-41
Number of pages6
JournalLife Sciences
Volume134
Issue number1
DOIs
Publication statusPublished - 2015 Jan 1
Externally publishedYes

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Keywords

  • Antioxidant
  • Cysteine-34
  • Genetic variants
  • Human serum albumin
  • Methionine
  • Recombinant mutants

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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