Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal rieske-type ferredoxin

Toshio Iwasaki, Risako Fukazawa, Yoshiharu Miyajima-Nakano, Amgalanbaatar Baldansuren, Shinichi Matsushita, Myat T. Lin, Robert B Gennis, Kazuya Hasegawa, Takashi Kumasaka, Sergei Dikanov

Research output: Contribution to journalArticle

Abstract

The electronic structure and geometry of redox-active metal cofactors in proteins are tuned by the pattern of hydrogen bonding with the backbone peptide matrix. In this study we developed a method for selective amino acid labeling of a hyperthermophilic archaeal metalloprotein with engineered Escherichia coli auxotroph strains, and we applied this to resolve the hydrogen bond interactions with the reduced Rieske-type [2Fe-2S] cluster by two-dimensional pulsed electron spin resonance technique. Because deep electron spin-echo envelope modulation of two histidine 14Nδ ligands of the cluster decreased non-coordinating 15N signal intensities via the cross-suppression effect, an inverse labeling strategy was employed in which 14N amino acid-labeled archaeal Rieske-type ferredoxin samples were examined in an 15N-protein background. This has directly identified Lys45 Nα as providing the major pathway for the transfer of unpaired electron spin density from the reduced cluster by a "through-bond" mechanism. All other backbone peptide nitrogens interact more weakly with the reduced cluster. The extension of this approach will allow visualizing the three-dimensional landscape of preferred pathways for the transfer of unpaired spin density from a paramagnetic metal center onto the protein frame, and will discriminate specific interactions by a "through-bond" mechanism from interactions which are "through-space" in various metalloproteins.

Original languageEnglish (US)
Pages (from-to)19731-19738
Number of pages8
JournalJournal of the American Chemical Society
Volume134
Issue number48
DOIs
StatePublished - Dec 5 2012

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Isotope Labeling
Ferredoxins
Dissection
dissection
Sulfur
Isotopes
Metalloproteins
Labeling
Hydrogen
Hydrogen bonds
Iron
sulfur
hydrogen
isotope
Proteins
iron
peptide
Peptides
protein
Amino acids

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal rieske-type ferredoxin. / Iwasaki, Toshio; Fukazawa, Risako; Miyajima-Nakano, Yoshiharu; Baldansuren, Amgalanbaatar; Matsushita, Shinichi; Lin, Myat T.; Gennis, Robert B; Hasegawa, Kazuya; Kumasaka, Takashi; Dikanov, Sergei.

In: Journal of the American Chemical Society, Vol. 134, No. 48, 05.12.2012, p. 19731-19738.

Research output: Contribution to journalArticle

Iwasaki, Toshio ; Fukazawa, Risako ; Miyajima-Nakano, Yoshiharu ; Baldansuren, Amgalanbaatar ; Matsushita, Shinichi ; Lin, Myat T. ; Gennis, Robert B ; Hasegawa, Kazuya ; Kumasaka, Takashi ; Dikanov, Sergei. / Dissection of hydrogen bond interaction network around an iron-sulfur cluster by site-specific isotope labeling of hyperthermophilic archaeal rieske-type ferredoxin. In: Journal of the American Chemical Society. 2012 ; Vol. 134, No. 48. pp. 19731-19738.
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