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Cited by 148Metalloenzymes and iron-sulfur proteinsMitochondrial Function and PathologyEnzyme Structure and Function

Human Mitochondrial Ferredoxin 1 (FDX1) and Ferredoxin 2 (FDX2) Both Bind Cysteine Desulfurase and Donate Electrons for Iron–Sulfur Cluster Biosynthesis

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Cai, Kai, Tonelli, Marco, Frederick, Ronnie O., Markley, John L.·2017·Biochemistry
DOI 10.1021/acs.biochem.6b00447PMC PMC5267338
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Abstract

Ferredoxins play an important role as an electron donor in iron–sulfur (Fe–S) cluster biosynthesis. Two ferredoxins, human mitochondrial ferredoxin 1 (FDX1) and human mitochondrial ferredoxin 2 (FDX2), are present in the matrix of human mitochondria. Conflicting results have been reported regarding their respective function in mitochondrial iron–sulfur cluster biogenesis. We report here biophysical studies of the interaction of these two ferredoxins with other proteins involved in mitochondrial iron–sulfur cluster assembly. Results from nuclear magnetic resonance spectroscopy show that both FDX1 and FDX2 (in both their reduced and oxidized states) interact with the protein complex responsible for cluster assembly, which contains cysteine desulfurase (NFS1), ISD11 (also known as LYRM4), and acyl carrier protein (Acp). In all cases, ferredoxin residues close to the Fe–S cluster are involved in the interaction with this complex. Isothermal titration calorimetry results showed that FDX2 binds more tightly to the cysteine desulfurase complex than FDX1 does. The reduced form of each ferredoxin became oxidized in the presence of the cysteine desulfurase complex when l-cysteine was added, leading to its conversion to l-alanine and the generation of sulfide. In an in vitro reaction, the reduced form of each ferredoxin was found to support Fe–S cluster assembly on ISCU; the rate of cluster assembly was faster with FDX2 than with FDX1. Taken together, these results show that both FDX1 and FDX2 can function in Fe–S cluster assembly in vitro.

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Authors
Cai, K., Tonelli, M., Frederick, R. O., & Markley, J. L.
Affiliations
University of Wisconsin–Madison (US)
Corresponding author
Markley, John L.
ORCID
Venue
Biochemistry
Publisher
American Chemical Society (ACS)
Published
2016-12-21
Volume
56
Issue
3
Pages
487-499
ISSN
  • 0006-2960 (ISSN-L)
  • 1520-4995
DOI
10.1021/acs.biochem.6b00447
PMC
PMC5267338
OpenAlex
W2565605212
Cited by
148
References
2
Citations by year
2026: 12 · 2025: 24 · 2024: 22 · 2023: 17 · 2022: 21 · 2021: 20 · 2020: 8 · 2019: 5 · 2018: 10 · 2017: 9
FWCI
2.17
Type
journal article
Language
en
Open access
Licence
Locations
  • Biochemistry (publishedVersion, other-oa)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute of General Medical Sciences (U01 GM94622)
Topics
  • Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
Keywords
FDX1, cysteine desulfurase, NFS1, acyl carrier protein, IscU, electron transfer, protein-protein interaction, sulfide generation, human mitochondria
MeSH terms
Frataxin, Acyl Carrier Protein, Amino Acid Sequence, Animals, Cysteine, Electrons, Escherichia coli, Ferredoxins, Gene Expression Regulation, Humans, Iron, Iron-Sulfur Proteins, Mitochondria, Models, Molecular, Oxidation-Reduction, Recombinant Proteins, Saccharomyces cerevisiae, Carbon-Sulfur Lyases, Sulfur, Sequence Alignment, Protein Structure, Secondary, Iron-Binding Proteins, Iron-Regulatory Proteins

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