The CcmC:Heme:CcmE Complex in Heme Trafficking and Cytochrome c Biosynthesis
Open accessAbstract
A superfamily of integral membrane proteins is characterized by a conserved tryptophan-rich region (called the WWD domain) in an external loop at the inner membrane surface. The three major members of this family (CcmC, CcmF, and CcsBA) are each involved in cytochrome c biosynthesis, yet the function of the WWD domain is unknown. It has been hypothesized that the WWD domain binds heme to present it to an acceptor protein (apoCcmE for CcmC or apocytochrome c for CcmF and CcsBA) such that the heme vinyl group(s) covalently attaches to the acceptors. Alternative proposals suggest that the WWD domain interacts directly with the acceptor protein (e.g., apoCcmE for CcmC). Here, it is shown that CcmC is only trapped with heme when its cognate acceptor protein CcmE is present. It is demonstrated that CcmE only interacts stably with CcmC when heme is present; thus, specific residues in each protein provide sites of interaction with heme to form this very stable complex. For the first time, evidence that the external WWD domain of CcmC interacts directly with heme is presented. Single and multiple substitutions of completely conserved residues in the WWD domain of CcmC alter the spectral properties of heme in the stable CcmC:heme:CcmE complexes. Moreover, some mutations reduce the binding of heme up to 100%. It is likely that endogenously synthesized heme enters the external WWD domain of CcmC either via a channel within this six-transmembrane-spanning protein or from the membrane. The data suggest that a specific heme channel (i.e., heme binding site within membrane spanning helices) is not present in CcmC, in contrast to the CcsBA protein. We discuss the likelihood that it is not important to protect the heme via trafficking in CcmC whereas it is critical in CcsBA.
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The pore may allow the access of heme from the outer leaflet, but no channel for potential heme trafficking is present within CcmC, as previously speculated.
“The pore may allow the access of heme from the outer leaflet, but no channel for potential heme trafficking is present within CcmC, as previously speculated48.”
From AF2Complex predicts direct physical interactions in multimeric proteins with deep learning (2022) · cites this paper as [48]
The interactions involve Trp114 and Trp119 of a tryptophan-rich loop (WWD domain), sitting at one edge of the binding pocket of the heme.
“The interactions involve Trp114 and Trp119 of a tryptophan-rich loop (WWD domain48), sitting at one edge of the binding pocket of the heme (Fig. 6c).”
From AF2Complex predicts direct physical interactions in multimeric proteins with deep learning (2022) · cites this paper as [48]
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- Authors
- Richard-Fogal, C., & Kranz, R. G.
- Affiliations
- Washington University in St. Louis (US)
- Corresponding author
- Kranz, Robert G.
- ORCID
- Kranz, Robert G.
- Venue
- Journal of Molecular Biology
- Publisher
- Elsevier BV
- Published
- 2010-08
- Volume
- 401
- Issue
- 3
- Pages
- 350-362
- ISSN
- 0022-2836 (ISSN-L)
- PMC
- PMC3580838
- OpenAlex
- W2034243710
- Cited by
- 53
- References
- 53
- Citations by year
- 2026: 1 · 2025: 4 · 2023: 4 · 2022: 3 · 2021: 5 · 2020: 1 · 2018: 5 · 2017: 2 · 2016: 4 · 2015: 1 · 2014: 3 · 2013: 7 · 2012: 5
- FWCI
- 2.99
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (GM47909)
- Topics
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Metalloenzymes and iron-sulfur proteins
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Photosynthetic Processes and Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Keywords
- CCMC, heme trafficking, heme binding, heme-protein complex, heme binding site, protein-protein interaction
- MeSH terms
- Bacterial Outer Membrane Proteins, Bacterial Proteins, Binding Sites, Biological Transport, Heme, Hemeproteins, Membrane Proteins, Protein Binding, Amino Acid Substitution, Protein Transport, Escherichia coli Proteins, Cytochromes c, Multiprotein Complexes
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