A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer

Anindya Roy, Dayn Joseph Sommer, Robert Arthur Schmitz, Chelsea Lynn Brown, Devens Gust, Andrei Astashkin, Giovanna Ghirlanda

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

[Fe-S] clusters, natures modular electron transfer units, are often arranged in chains that support long-range electron transfer. Despite considerable interest, the design of biomimetic artificial systems emulating multicluster-binding proteins, with the final goal of integrating them in man-made oxidoreductases, remains elusive. Here, we report a novel bis-[4Fe-4S] cluster binding protein, DSD-Fdm, in which the two clusters are positioned within a distance of 12 Å, compatible with the electronic coupling necessary for efficient electron transfer. The design exploits the structural repeat of coiled coils as well as the symmetry of the starting scaffold, a homodimeric helical protein (DSD). In total, eight hydrophobic residues in the core of DSD were replaced by eight cysteine residues that serve as ligands to the [4Fe-4S] clusters. Incorporation of two [4Fe-4S] clusters proceeds with high yield. The two [4Fe-4S] clusters are located in the hydrophobic core of the helical bundle as characterized by various biophysical techniques. The secondary structure of the apo and holo proteins is conserved; further, the incorporation of clusters results in stabilization of the protein with respect to chemical denaturation. Most importantly, this de novo designed protein can mimic the function of natural ferredoxins: we show here that reduced DSD-Fdm transfers electrons to cytochrome c, thus generating the reduced cyt c stoichiometrically.

Original languageEnglish (US)
Pages (from-to)17343-17349
Number of pages7
JournalJournal of the American Chemical Society
Volume136
Issue number49
DOIs
StatePublished - Dec 10 2014

Fingerprint

Ferredoxins
Electrons
Proteins
Carrier Proteins
Biomimetics
Denaturation
Cytochromes c
Scaffolds
Cysteine
Scaffolds (biology)
Oxidoreductases
Stabilization
Ligands

ASJC Scopus subject areas

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

Cite this

Roy, A., Sommer, D. J., Schmitz, R. A., Brown, C. L., Gust, D., Astashkin, A., & Ghirlanda, G. (2014). A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer. Journal of the American Chemical Society, 136(49), 17343-17349. https://doi.org/10.1021/ja510621e

A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer. / Roy, Anindya; Sommer, Dayn Joseph; Schmitz, Robert Arthur; Brown, Chelsea Lynn; Gust, Devens; Astashkin, Andrei; Ghirlanda, Giovanna.

In: Journal of the American Chemical Society, Vol. 136, No. 49, 10.12.2014, p. 17343-17349.

Research output: Contribution to journalArticle

Roy, A, Sommer, DJ, Schmitz, RA, Brown, CL, Gust, D, Astashkin, A & Ghirlanda, G 2014, 'A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer', Journal of the American Chemical Society, vol. 136, no. 49, pp. 17343-17349. https://doi.org/10.1021/ja510621e
Roy A, Sommer DJ, Schmitz RA, Brown CL, Gust D, Astashkin A et al. A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer. Journal of the American Chemical Society. 2014 Dec 10;136(49):17343-17349. https://doi.org/10.1021/ja510621e
Roy, Anindya ; Sommer, Dayn Joseph ; Schmitz, Robert Arthur ; Brown, Chelsea Lynn ; Gust, Devens ; Astashkin, Andrei ; Ghirlanda, Giovanna. / A de novo designed 2[4Fe-4S] ferredoxin mimic mediates electron transfer. In: Journal of the American Chemical Society. 2014 ; Vol. 136, No. 49. pp. 17343-17349.
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