The CcmFH complex is the system I holocytochrome c synthetase: engineering cytochrome c maturation independent of CcmABCDE
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Summary: Cytochrome c maturation (ccm) in many bacteria, archaea and plant mitochondria requires eight membrane proteins, CcmABCDEFGH, called system I. This pathway delivers and attaches haem covalently to two cysteines (of Cys‐X xx‐X xx‐Cys‐His) in the cytochrome c. All models propose that CcmFH facilitates covalent attachment of haem to the apocytochrome; namely, that it is the synthetase. However, holocytochrome c synthetase activity has not been directly demonstrated for CcmFH. We report formation of holocytochromes c by CcmFH and CcmG, a periplasmic thioredoxin, independent of CcmABCDE (we term this activity CcmFGH‐only). Cytochrome c produced in the absence of CcmABCDE is indistinguishable from cytochrome c produced by the full system I, with a cleaved signal sequence and two covalent bonds to haem. We engineered increased cytochrome c production by CcmFGH‐only, with yields approaching those from the full system I. Three conserved histidines in CcmF (TM‐His 1, TM‐His 2 and P‐His 1) are required for activity, as are the conserved cysteine pairs in CcmG and CcmH. Our findings establish that CcmFH is the system I holocytochrome c synthetase. Although we discuss why this engineering would likely not replace the need for CcmABCDE in nature, these results provide unique mechanistic and evolutionary insights into cytochrome c biosynthesis.
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CcmFGH covalently attaches the heme to nascent cytochrome c-type proteins.
“Subsequently, CcmFGH covalently attaches the heme to nascent cytochrome c-type proteins45,46.”
From AF2Complex predicts direct physical interactions in multimeric proteins with deep learning (2022) · cites this paper as [45]
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AF2Complex predicts direct physical interactions in multimeric proteins with deep learning
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- Authors
- San Francisco, B., Sutherland, M. C., & Kranz, R. G.
- Affiliations
- Department of Biology Washington University in St. Louis St. Louis MO 63130 USA
- Washington University in St. Louis (US)
- Corresponding author
- Kranz, Robert G.
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 2014-01-27
- Volume
- 91
- Issue
- 5
- Pages
- 996-1008
- ISSN
- 0950-382X (ISSN-L)
- 1365-2958
- PMC
- PMC3959880
- OpenAlex
- W1859783629
- Cited by
- 29
- References
- 58
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 1 · 2023: 3 · 2022: 2 · 2021: 5 · 2020: 2 · 2019: 2 · 2018: 2 · 2017: 1 · 2016: 3 · 2015: 3
- FWCI
- 0.77
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Microbiology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (GM103422, R01 GM47909, GM47909)
- National Institute of General Medical Sciences (GM103422, R01 GM47909, R01GM047909, P41GM103422)
- Topics
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- cytochrome c maturation, conserved histidine, cysteine pair, protein engineering
- MeSH terms
- Amino Acid Sequence, Cysteine, Escherichia coli, Heme, Histidine, Lyases, Molecular Sequence Data, Protein Engineering, Conserved Sequence, Amino Acid Substitution, Escherichia coli Proteins, Cytochromes c, Multiprotein Complexes
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