6QJV

R2-like ligand-binding oxidase E69D mutant with aerobically reconstituted Mn/Fe cofactor

Summary for 6QJV

• Entry DOI: 10.2210/pdb6qjv/pdb
• Related: 4HR0 4HR4 4HR5 4XB9 4XBV 4XBW 5DCO 5DCR 5DCS 5EKB 5OMJ 5OMK 6F65 6F6B 6F6C 6F6E 6F6F 6F6G 6F6H 6F6K 6F6L 6F6M 6I90 6I92 6I93 6I94 6I95 6QK0 6QK1 6QK2
• 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 (strain HTA426)
• Total number of polymer chains: 1
• Total formula weight: 37370.93

Authors

Griese, J.J.,Hogbom, M. (deposition date: 2019-01-25, release date: 2020-02-12, 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 (1.905 Å)