In vitro reconstitution reveals major differences between human and bacterial cytochrome c synthases
| Author(s) | Sutherland, Molly C. | |
| Author(s) | Mendez, Deanna L. | |
| Author(s) | Babbitt, Shalon E. | |
| Author(s) | Tillman, Dustin E. | |
| Author(s) | Melnikov, Olga | |
| Author(s) | Tran, Nathan L. | |
| Author(s) | Prizant, Noah T. | |
| Author(s) | Collier, Andrea L. | |
| Author(s) | Kranz, Robert G. | |
| Date Accessioned | 2021-12-22T19:24:59Z | |
| Date Available | 2021-12-22T19:24:59Z | |
| Publication Date | 2021-05-11 | |
| Description | This article was originally published in eLife. The version of record is available at: https://doi.org/10.7554/eLife.64891 | en_US |
| Abstract | Cytochromes c are ubiquitous heme proteins in mitochondria and bacteria, all possessing a CXXCH (CysXxxXxxCysHis) motif with covalently attached heme. We describe the first in vitro reconstitution of cytochrome c biogenesis using purified mitochondrial (HCCS) and bacterial (CcsBA) cytochrome c synthases. We employ apocytochrome c and peptide analogs containing CXXCH as substrates, examining recognition determinants, thioether attachment, and subsequent release and folding of cytochrome c. Peptide analogs reveal very different recognition requirements between HCCS and CcsBA. For HCCS, a minimal 16-mer peptide is required, comprised of CXXCH and adjacent alpha helix 1, yet neither thiol is critical for recognition. For bacterial CcsBA, both thiols and histidine are required, but not alpha helix 1. Heme attached peptide analogs are not released from the HCCS active site; thus, folding is important in the release mechanism. Peptide analogs behave as inhibitors of cytochrome c biogenesis, paving the way for targeted control. | en_US |
| Sponsor | Funder: National Institutes of Health Grant reference number: (R01 GM47909) Author: Robert G Kranz The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. | en_US |
| Citation | Sutherland, M. C., Mendez, D. L., Babbitt, S. E., Tillman, D. E., Melnikov, O., Tran, N. L., Prizant, N. T., Collier, A. L., & Kranz, R. G. (2021). In vitro reconstitution reveals major differences between human and bacterial cytochrome c synthases. ELife, 10, e64891. https://doi.org/10.7554/eLife.64891 | en_US |
| ISSN | 2050-084X | |
| URL | https://udspace.udel.edu/handle/19716/29912 | |
| Language | en_US | en_US |
| Publisher | eLife | en_US |
| Title | In vitro reconstitution reveals major differences between human and bacterial cytochrome c synthases | en_US |
| Type | Article | en_US |
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