85 publications

85 publications

A Cell-Penetrating Artificial Metalloenzyme Regulates a Gene Switch in a Designer Mammalian Cell

Fussenegger, M.; Matile, S.; Ward, T. R.

Nat. Commun., 2018, 10.1038/s41467-018-04440-0

Complementing enzymes in their native environment with either homogeneous or heterogeneous catalysts is challenging due to the sea of functionalities present within a cell. To supplement these efforts, artificial metalloenzymes are drawing attention as they combine attractive features of both homogeneous catalysts and enzymes. Herein we show that such hybrid catalysts consisting of a metal cofactor, a cell-penetrating module, and a protein scaffold are taken up into HEK-293T cells where they catalyze the uncaging of a hormone. This bioorthogonal reaction causes the upregulation of a gene circuit, which in turn leads to the expression of a nanoluc-luciferase. Relying on the biotin–streptavidin technology, variation of the biotinylated ruthenium complex: the biotinylated cell-penetrating poly(disulfide) ratio can be combined with point mutations on streptavidin to optimize the catalytic uncaging of an allyl-carbamate-protected thyroid hormone triiodothyronine. These results demonstrate that artificial metalloenzymes offer highly modular tools to perform bioorthogonal catalysis in live HEK cells.


Metal: Ru
Ligand type: Cp; Quinoline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Deallylation
Max TON: 33
ee: ---
PDB: ---
Notes: ---

Achiral Cyclopentadienone Iron Tricarbonyl Complexes Embedded in Streptavidin: An Access to Artificial Iron Hydrogenases and Application in Asymmetric Hydrogenation

Renaud, J.-L.; Ward, T. R.

Catal. Lett., 2016, 10.1007/s10562-015-1681-6

We report on the synthesis of biotinylated (cyclopentadienone)iron tricarbonyl complexes, the in situ generation of the corresponding streptavidin conjugates and their application in asymmetric hydrogenation of imines and ketones.


Metal: Fe
Ligand type: CO; Cyclopentadienone
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical
Reaction: Hydrogenation
Max TON: 20
ee: 34
PDB: ---
Notes: ---

A Dual Anchoring Strategy for the Localization and Activation of Artificial Metalloenzymes Based on the Biotin−Streptavidin Technology

Ward, T. R.

J. Am. Chem. Soc., 2013, 10.1021/ja309974s

Artificial metalloenzymes result from anchoring an active catalyst within a protein environment. Toward this goal, various localization strategies have been pursued: covalent, supramolecular, or dative anchoring. Herein we show that introduction of a suitably positioned histidine residue contributes to firmly anchor, via a dative bond, a biotinylated rhodium piano stool complex within streptavidin. The in silico design of the artificial metalloenzyme was confirmed by X-ray crystallography. The resulting artificial metalloenzyme displays significantly improved catalytic performance, both in terms of activity and selectivity in the transfer hydrogenation of imines. Depending on the position of the histidine residue, both enantiomers of the salsolidine product can be obtained.


Metal: Ir
Ligand type: Amino acid; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 14
ee: 11
PDB: ---
Notes: ---

Metal: Rh
Ligand type: Amino acid; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 100
ee: 79
PDB: ---
Notes: ---

An Artificial Imine Reductase Based on the Ribonuclease S Scaffold

Ward, T. R.

ChemCatChem, 2014, 10.1002/cctc.201300995


Metal: Ir
Ligand type: Amino acid; Cp*
Host protein: Ribonuclease S
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 4
ee: 18
PDB: ---
Notes: ---

An Artificial Metalloenzyme for Carbene Transfer Based on a Biotinylated Dirhodium Anchored Within Streptavidin

Ward, T. R.

Cat. Sci. Technol., 2018, 10.1039/C8CY00646F


Metal: Rh
Ligand type: Carboxylate
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Cyclopropanation
Max TON: ~60
ee: ---
PDB: ---
Notes: Cyclopropanation reaction was also performed in the E. coli periplasm.

Metal: Rh
Ligand type: Carboxylate
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: C-H insertion
Max TON: ~60
ee: ---
PDB: ---
Notes: ---

An Artificial Metalloenzyme for Olefin Metathesis

Hilvert, D.; Ward, T. R.

Chem. Commun., 2011, 10.1039/c1cc15005g

A Grubbs–Hoveyda type olefin metathesis catalyst, equipped with an electrophilic bromoacetamide group, was used to modify a cysteine-containing variant of a small heat shock protein from Methanocaldococcus jannaschii. The resulting artificial metalloenzyme was found to be active under acidic conditions in a benchmark ring closing metathesis reaction.


Metal: Ru
Ligand type: Carbene
Anchoring strategy: Covalent
Optimization: ---
Reaction: Olefin metathesis
Max TON: 25
ee: ---
PDB: ---
Notes: RCM

An Enantioselective Artificial Suzukiase Based on the Biotin–Streptavidin Technology

Ward, T. R.

Chem. Sci., 2015, 10.1039/c5sc03116h


Metal: Pd
Ligand type: Allyl; Phosphine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 88
ee: 80
PDB: ---
Notes: ---

Metal: Pd
Ligand type: Allyl; Carbene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 5
ee: ---
PDB: ---
Notes: ---

An NAD(P)H-Dependent Artificial Transfer Hydrogenase for Multienzymatic Cascades

Ward, T. R.

J. Am. Chem. Soc., 2016, 10.1021/jacs.6b02470


Metal: Ir
Ligand type: Cp*; Phenanthroline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: >999
ee: >99
PDB: ---
Notes: ---

Aqueous Olefin Metathesis: Recent Developments and Applications

Review

Ward, T. R.

Beilstein J. Org. Chem., 2019, 10.3762/bjoc.15.39


Notes: ---

Aqueous Oxidation of Alcohols Catalyzed by Artificial Metalloenzymes Based on the Biotin–Avidin Technology

Ward, T. R.

J. Organomet. Chem., 2005, 10.1016/j.jorganchem.2005.02.001


Metal: Ru
Ligand type: Amino-sulfonamide; Benzene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Alcohol oxidation
Max TON: 200
ee: ---
PDB: ---
Notes: ---

Metal: Ru
Ligand type: Amino-sulfonamide; Benzene
Host protein: Avidin (Av)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Alcohol oxidation
Max TON: 230
ee: ---
PDB: ---
Notes: ---

Metal: Ru
Ligand type: Bipyridine; C6Me6
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Alcohol oxidation
Max TON: 173
ee: ---
PDB: ---
Notes: ---

Metal: Rh
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Alcohol oxidation
Max TON: 7.5
ee: ---
PDB: ---
Notes: ---

Metal: Ir
Ligand type: Bipyridine; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Alcohol oxidation
Max TON: 30
ee: ---
PDB: ---
Notes: ---

Artificial Metalloenzyme for Enantioselective Sulfoxidation Based on Vanadyl-Loaded Streptavidin

Ward, T. R.

J. Am. Chem. Soc., 2008, 10.1021/ja8017219


Metal: V
Ligand type: Water
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Sulfoxidation
Max TON: 27
ee: 93
PDB: ---
Notes: ---

Artificial Metalloenzymes

Review

Ward, T. R.

Effects of Nanoconfinement on Catalysis, 2017, 10.1007/978-3-319-50207-6_3


Notes: Book chapter

Artificial Metalloenzymes as Selective Catalysts in Aqueous Media

Review

Ward, T. R.

Coord. Chem. Rev., 2008, 10.1016/j.ccr.2007.09.016


Notes: ---

Artificial Metalloenzymes Based on Biotin-Avidin Technology for the Enantioselective Reduction of Ketones by Transfer Hydrogenation

Ward, T. R.

Proc. Natl. Acad. Sci. U. S. A., 2005, 10.1073/pnas.0409684102


Metal: Ru
Ligand type: Amino-sulfonamide; P-cymene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 92
ee: 94
PDB: ---
Notes: ---

Metal: Ru
Ligand type: Amino-sulfonamide; Benzene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 30
ee: 63
PDB: ---
Notes: ---

Artificial Metalloenzymes Based on the Biotin-Avidin Technology: Enantioselective Catalysis and Beyond

Review

Ward, T. R.

Acc. Chem. Res., 2011, 10.1021/ar100099u


Notes: ---

Artificial Metalloenzymes Based on the Biotin-Streptavidin Technology: Challenges and Opportunities

Review

Ward, T. R.

Acc. Chem. Res., 2016, 10.1021/acs.accounts.6b00235


Notes: ---

Artificial Metalloenzymes Based on the Biotin–Streptavidin Technology: Enzymatic Cascades and Directed Evolution

Review

Ward, T. R.

Acc. Chem. Res., 2019, 10.1021/acs.accounts.8b00618


Notes: ---

Artificial Metalloenzymes: Combining the Best Features of Homogeneous and Enzymatic Catalysis

Review

Ward, T. R.

Synlett, 2009, 10.1055/s-0029-1218305


Notes: ---

Artificial Metalloenzymes: Enantioselective Catalysis and Beyond

Review

Ward, T. R.

Chimia, 2010, 10.2533/chimia.2010.846


Notes: ---

Artificial Metalloenzymes for Asymmetric Allylic Alkylation on the Basis of the Biotin–Avidin Technology

Ward, T. R.

Angew. Chem., Int. Ed., 2008, 10.1002/anie.200703159


Metal: Pd
Ligand type: Phosphine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Allylic alkylation
Max TON: 10
ee: 93
PDB: ---
Notes: ---

Artificial Metalloenzymes for Enantioselective Catalysis Based on Biotin-Avidin

Ward, T. R.

J. Am. Chem. Soc., 2003, 10.1021/ja035545i


Metal: Rh
Ligand type: Phosphine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Hydrogenation
Max TON: ---
ee: 96
PDB: ---
Notes: ---

Artificial Metalloenzymes for Enantioselective Catalysis Based on the Biotin-Avidin Technology

Review

Ward, T. R.

Top. Organomet. Chem., 2009, 10.1007/3418_2008_3


Notes: Book chapter

Artificial Metalloenzymes for Enantioselective Catalysis Based on the Biotin-Avidin Technology

Review

Ward, T. R.

Chimia, 2008, 10.2533/chimia.2008.956


Notes: ---

Artificial Metalloenzymes for Enantioselective Catalysis Based on the Noncovalent Incorporation of Organometallic Moieties in a Host Protein

Review

Ward, T. R.

Chem. - Eur. J., 2005, 10.1002/chem.200401232


Notes: ---

Artificial Metalloenzymes for Enantioselective Catalysis: Recent Advances

Review

Ward, T. R.

ChemBioChem, 2006, 10.1002/cbic.200600264


Notes: ---

Artificial Metalloenzymes for Enantioselective Catalysis: The Phenomenon of Protein Accelerated Catalysis

Ward, T. R.

J. Organomet. Chem., 2004, 10.1016/j.jorganchem.2004.09.032


Metal: Rh
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical
Reaction: Hydrogenation
Max TON: ---
ee: 94
PDB: ---
Notes: Reduction of acetamidoacrylic acid. 3.0-fold protein acceleration.

Metal: Rh
Host protein: Avidin (Av)
Anchoring strategy: Supramolecular
Optimization: Chemical
Reaction: Hydrogenation
Max TON: ---
ee: 39
PDB: ---
Notes: Reduction of acetamidoacrylic acid. 12.0-fold protein acceleration.

Artificial Metalloenzymes for Olefin Metathesis Based on the Biotin-(Strept)Avidin Technology

Ward, T. R.

Chem. Commun., 2011, 10.1039/c1cc15004a


Metal: Ru
Ligand type: Carbene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical
Reaction: Olefin metathesis
Max TON: 14
ee: ---
PDB: ---
Notes: RCM

Metal: Ru
Ligand type: Carbene
Host protein: Avidin (Av)
Anchoring strategy: Supramolecular
Optimization: Chemical
Reaction: Olefin metathesis
Max TON: 19
ee: ---
PDB: ---
Notes: RCM

Artificial Metalloenzymes for the Diastereoselective Reduction of NAD+ to NAD2H

Ward, T. R.

Org. Biomol. Chem., 2015, 10.1039/c4ob02071e


Metal: Ir
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: ---
Max TON: ---
ee: ---
PDB: ---
Notes: ---

Artificial Metalloenzymes: Proteins as Hosts for Enantioselective Catalysis

Review

Ward, T. R.

Chem. Soc. Rev., 2005, 10.1039/b314695m


Notes: ---

Artificial Metalloenzymes: Reaction Scope and Optimization Strategies

Review

Lewis, J. C.; Ward, T. R.

Chem. Rev., 2018, 10.1021/acs.chemrev.7b00014


Notes: ---