6U97

Structure of OmcF_H47I mutant

Summary for 6U97

• Entry DOI: 10.2210/pdb6u97/pdb
• Descriptor: Lipoprotein cytochrome c, 1 heme-binding site, SULFATE ION, PROTOPORPHYRIN IX CONTAINING FE, ... (4 entities in total)
• Functional Keywords: omcf, redox-bohr effect, electron transport
• Biological source: Geobacter sulfurreducens (strain ATCC 51573 / DSM 12127 / PCA)
• Total number of polymer chains: 1
• Total formula weight: 9140.15

Authors

Pokkuluri, P.R. (deposition date: 2019-09-06, release date: 2020-02-19, Last modification date: 2024-11-13)

Primary citation

Teixeira, L.R.,Cordas, C.M.,Fonseca, M.P.,Duke, N.E.C.,Pokkuluri, P.R.,Salgueiro, C.A.
Modulation of the Redox Potential and Electron/Proton Transfer Mechanisms in the Outer Membrane Cytochrome OmcF FromGeobacter sulfurreducens.
Front Microbiol, 10:2941-2941, 2019

PubMed Abstract: The monoheme outer membrane cytochrome F (OmcF) from plays an important role in Fe(III) reduction and electric current production. The electrochemical characterization of this cytochrome has shown that its redox potential is modulated by the solution pH (redox-Bohr effect) endowing the protein with the necessary properties to couple electron and proton transfer in the physiological range. The analysis of the OmcF structures in the reduced and oxidized states showed that with the exception of the side chain of histidine 47 (His), all other residues with protonatable side chains are distant from the heme iron and, therefore, are unlikely to affect the redox potential of the protein. The protonatable site at the imidazole ring of His is in the close proximity to the heme and, therefore, this residue was suggested as the redox-Bohr center. In the present work, we tested this hypothesis by replacing the His with non-protonatable residues (isoleucine - OmcFH47I and phenylalanine - OmcFH47F). The structure of the mutant OmcFH47I was determined by X-ray crystallography to 1.13 Å resolution and showed only minimal changes at the site of the mutation. Both mutants were N-labeled and their overall folding was confirmed to be the same as the wild-type by NMR spectroscopy. The pH dependence of the redox potential of the mutants was measured by cyclic voltammetry. Compared to the wild-type protein, the magnitude of the redox-Bohr effect in the mutants was smaller, but not fully abolished, confirming the role of His on the pH modulation of OmcF's redox potential. However, the pH effect on the heme substituents' NMR chemical shifts suggested that the heme propionate P also contributes to the overall redox-Bohr effect in OmcF. In physiological terms, the contribution of two independent acid-base centers to the observed redox-Bohr effect confers OmcF a higher versatility to environmental changes by coupling electron/proton transfer within a wider pH range.

PubMed: 32010071
DOI: 10.3389/fmicb.2019.02941

Experimental method

X-RAY DIFFRACTION (1.13 Å)