Spectroscopic and Electrochemical Characterization of the Mycofactocin Biosynthetic Protein, MftC, Provides Insight into Its Redox Flipping Mechanism

Mycofactocin is a putative redox cofactor and is classified as a ribosomally synthesized and post-translationally modified peptide (RiPP). Some RiPP natural products, including mycofactocin, rely on a radical S-adenosylmethionine (RS, SAM) protein to modify the precursor peptide. Mycofactocin matura...

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Veröffentlicht in:Biochemistry (Easton) 2019-02, Vol.58 (7), p.940-950
Hauptverfasser: Ayikpoe, Richard, Ngendahimana, Thacien, Langton, Michelle, Bonitatibus, Sheila, Walker, Lindsey M, Eaton, Sandra S, Eaton, Gareth R, Pandelia, Maria-Eirini, Elliott, Sean J, Latham, John A
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Sprache:eng
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Zusammenfassung:Mycofactocin is a putative redox cofactor and is classified as a ribosomally synthesized and post-translationally modified peptide (RiPP). Some RiPP natural products, including mycofactocin, rely on a radical S-adenosylmethionine (RS, SAM) protein to modify the precursor peptide. Mycofactocin maturase, MftC, is a unique RS protein that catalyzes the oxidative decarboxylation and C–C bond formation on the precursor peptide MftA. However, the number, chemical nature, and catalytic roles for the MftC [Fe–S] clusters remain unknown. Here, we report that MftC binds a RS [4Fe–4S] cluster and two auxiliary [4Fe–4S] clusters that are required for MftA modification. Furthermore, electron paramagnetic resonance spectra of MftC suggest that SAM and MftA affect the environments of the RS and Aux I cluster, whereas the Aux II cluster is unaffected by the substrates. Lastly, reduction potential assignments of individual [4Fe–4S] clusters by protein film voltammetry show that their potentials are within 100 mV of each other.
ISSN:0006-2960
1520-4995
DOI:10.1021/acs.biochem.8b01082