Controlled Ligand Exchange Between Ruthenium Organometallic Cofactor Precursors and a Naïve Protein Scaffold Generates Artificial Metalloenzymes Catalysing Transfer Hydrogenation.
George S BiggsOskar James KleinSarah L MaslenJ Mark SkehelTrevor J RutherfordStefan M V FreundFlorian HollfelderSally R BossPaul D BarkerPublished in: Angewandte Chemie (International ed. in English) (2021)
Many natural metalloenzymes assemble from proteins and biosynthesised complexes, generating potent catalysts by changing metal coordination. Here we adopt the same strategy to generate artificial metalloenzymes (ArMs) using ligand exchange to unmask catalytic activity. By systematically testing RuII (η6 -arene)(bipyridine) complexes designed to facilitate the displacement of functionalised bipyridines, we develop a fast and robust procedure for generating new enzymes via ligand exchange in a protein that has not evolved to bind such a complex. The resulting metal cofactors form peptidic coordination bonds but also retain a non-biological ligand. Tandem mass spectrometry and 19 F NMR spectroscopy were used to characterise the organometallic cofactors and identify the protein-derived ligands. By introduction of ruthenium cofactors into a 4-helical bundle, transfer hydrogenation catalysts were generated that displayed a 35-fold rate increase when compared to the respective small molecule reaction in solution.
Keyphrases
- small molecule
- tandem mass spectrometry
- protein protein
- ultra high performance liquid chromatography
- high performance liquid chromatography
- binding protein
- highly efficient
- liquid chromatography
- amino acid
- simultaneous determination
- gas chromatography
- high resolution
- minimally invasive
- mass spectrometry
- solid phase extraction
- transition metal
- metal organic framework
- solid state