6QK1

R2-like ligand-binding oxidase Y175F mutant with aerobically reconstituted Mn/Fe cofactor

Summary for 6QK1

• Entry DOI: 10.2210/pdb6qk1/pdb
• Descriptor: Ribonucleotide reductase small subunit, PALMITIC ACID, MANGANESE (III) ION, ... (6 entities in total)
• Functional Keywords: r2-like ligand-binding oxidase, mn/fe cofactor, ribonucleotide reductase r2 subunit fold, metalloprotein oxidoreductase, oxidoreductase
• Biological source: Geobacillus kaustophilus HTA426
• Total number of polymer chains: 1
• Total formula weight: 37368.95

Authors

Griese, J.J.,Hogbom, M. (deposition date: 2019-01-28, release date: 2020-02-19, Last modification date: 2024-01-24)

Primary citation

Kisgeropoulos, E.C.,Griese, J.J.,Smith, Z.R.,Branca, R.M.M.,Schneider, C.R.,Hogbom, M.,Shafaat, H.S.
Key Structural Motifs Balance Metal Binding and Oxidative Reactivity in a Heterobimetallic Mn/Fe Protein.
J.Am.Chem.Soc., 142:5338-5354, 2020

PubMed Abstract: Heterobimetallic Mn/Fe proteins represent a new cofactor paradigm in bioinorganic chemistry and pose countless outstanding questions. The assembly of the active site defies common chemical convention by contradicting the Irving-Williams series, while the scope of reactivity remains unexplored. In this work, the assembly and C-H bond activation process in the Mn/Fe R2-like ligand-binding oxidase (R2lox) protein is investigated using a suite of biophysical techniques, including time-resolved optical spectroscopy, global kinetic modeling, X-ray crystallography, electron paramagnetic resonance spectroscopy, protein electrochemistry, and mass spectrometry. Selective metal binding is found to be under thermodynamic control, with the binding sites within the apo-protein exhibiting greater Mn affinity than Fe affinity. The comprehensive analysis of structure and reactivity of wild-type R2lox and targeted primary and secondary sphere mutants indicate that the efficiency of C-H bond activation directly correlates with the Mn/Fe cofactor reduction potentials and is inversely related to divalent metal binding affinity. These findings suggest the R2lox active site is precisely tuned for achieving both selective heterobimetallic binding and high levels of reactivity and offer a mechanism to examine the means by which proteins achieve appropriate metal incorporation.

PubMed: 32062969
DOI: 10.1021/jacs.0c00333

Experimental method

X-RAY DIFFRACTION (1.754 Å)