| Title: |
Cyanophenylalanine as an infrared probe for iron–sulfur cluster redox state in multicenter metalloenzymes |
| Authors: |
Duan, Zehui; Wei, Jiaao; Carr, Stephen B.; Ramirez, Miguel; Evans, Rhiannon M.; Ash, Philip A.; Rodriguez-Macia, Patricia; Sachdeva, Amit; Vincent, Kylie A. |
| Publication Year: |
2025 |
| Collection: |
University of East Anglia: UEA Digital Repository |
| Description: |
The noncanonical amino acid, para-cyanophenylalanine (CNF), when incorporated into metalloproteins, functions as an infrared spectroscopic probe for the redox state of iron-sulfur clusters, offering a strategy for determining electron occupancy in the electron transport chains of complex metalloenzymes. A redshift of ≈1–2 cm−1 in the nitrile (NC) stretching frequency is observed, following reduction of spinach ferredoxin modified to contain CNF close to its [2Fe–2S] center, and this shift is reversed on re-oxidation. We extend this to CNF positioned near to the proximal [4Fe–4S] cluster of the [FeFe] hydrogenase from Desulfovibrio desulfuricans. In combination with a distal [4Fe–4S] cluster and the [4Fe–4S] cluster of the active site ‘H-cluster’ ([4Fe–4S]H), the proximal cluster forms an electron relay connecting the active site to the surface of the protein. Again, a reversible shift in wavenumber for CNF is observed, following cluster reduction in either apo-protein (containing the iron-sulfur clusters but lacking the active site) or holo-protein with intact active site, demonstrating the general applicability of this approach to studying complex metalloenzymes. |
| Document Type: |
article in journal/newspaper |
| File Description: |
application/pdf |
| Language: |
English |
| ISSN: |
1439-4227 |
| Relation: |
https://ueaeprints.uea.ac.uk/id/eprint/100428/1/Duan_etal_2025_ChemBioChem.pdf; Duan, Zehui, Wei, Jiaao, Carr, Stephen B., Ramirez, Miguel, Evans, Rhiannon M., Ash, Philip A., Rodriguez-Macia, Patricia, Sachdeva, Amit and Vincent, Kylie A. (2025) Cyanophenylalanine as an infrared probe for iron–sulfur cluster redox state in multicenter metalloenzymes. ChemBioChem, 26 (14). ISSN 1439-4227 |
| DOI: |
10.1002/cbic.202500251 |
| Availability: |
https://ueaeprints.uea.ac.uk/id/eprint/100428/; https://ueaeprints.uea.ac.uk/id/eprint/100428/1/Duan_etal_2025_ChemBioChem.pdf; https://doi.org/10.1002/cbic.202500251 |
| Rights: |
cc_by |
| Accession Number: |
edsbas.38802A31 |
| Database: |
BASE |