Proteins as diverse, efficient, and evolvable scaffolds for artificial metalloenzymes

Jeong, Woo Jae; Yu, Jaeseung; Song, Woon Ju

doi:10.1039/d0cc03137b

Cited by 33 publications

(27 citation statements)

References 150 publications

(161 reference statements)

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“…Thus, identifying a suitable scaffold protein is essential for improving ArMs. 4 Multiple scaffolds have been explored for ArMs, including heme proteins, 5 , 6 (strept)avidin, 7 human carbonic anhydrase, 8 glycosylated albumin, 9 lactococcal multidrug resistance regulator (LmrR), 10 an oligopeptidase, 11 FhuA, 12 and so on. 13 Increasing the number of viable scaffolds will eventually enable chemists to use ArMs as a plug-and-play strategy, in which several scaffolds can be screened to identify the best catalytic starting point.…”

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confidence: 99%

Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

2021

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Artificial metalloenzymes (ArMs) are created by embedding a synthetic metal catalyst into a protein scaffold. ArMs have the potential to merge the catalytic advantages of natural enzymes with the reaction scope of synthetic catalysts. The choice of the protein scaffold is of utmost importance to tune the activity of the ArM. Herein, we show the repurposing of HaloTag, a self-labeling protein widely used in chemical biology, to create an ArM scaffold for metathesis. This monomeric protein scaffold allows for covalent attachment of metathesis cofactors, and the resulting ArMs are capable of catalyzing ring-closing metathesis. Both chemical and genetic engineering were explored to determine the evolvability of the resulting ArM. Additionally, exploration of the substrate scope revealed a reaction with promising turnover numbers (>48) and conversion rates (>96%).

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confidence: 99%

Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

2021

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“…Cysteine residues have also been explored as a reactive handle for the selective anchoring of metal-coordinating ligands or complexes. As a result, different proteins scaffolds can be converted into semi-synthetic hybrid catalysts, known as artificial metalloenzymes [102]. Steroid Carrier Protein 2L mutant (A100C) was converted into a metalloenzyme through tethering of iron-binding nitrogen cofactor containing a maleimide linker [103].…”

Section: Modification At Cysteinesmentioning

confidence: 99%

Protein Modifications: From Chemoselective Probes to Novel Biocatalysts

2021

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Chemical reactions can be performed to covalently modify specific residues in proteins. When applied to native enzymes, these chemical modifications can greatly expand the available set of building blocks for the development of biocatalysts. Nucleophilic canonical amino acid sidechains are the most readily accessible targets for such endeavors. A rich history of attempts to design enhanced or novel enzymes, from various protein scaffolds, has paved the way for a rapidly developing field with growing scientific, industrial, and biomedical applications. A major challenge is to devise reactions that are compatible with native proteins and can selectively modify specific residues. Cysteine, lysine, N-terminus, and carboxylate residues comprise the most widespread naturally occurring targets for enzyme modifications. In this review, chemical methods for selective modification of enzymes will be discussed, alongside with examples of reported applications. We aim to highlight the potential of such strategies to enhance enzyme function and create novel semisynthetic biocatalysts, as well as provide a perspective in a fast-evolving topic.

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“…Recent advances dramatically expand the range of chemical moieties available on the side chains through incorporation of noncanonical amino acids (Alcala-Torano et al, 2016;Burke et al, 2019;Zhou and Roelfes, 2020). Further, proteins can be optimized by directed evolution coupled with high-thrughput screening to identify favorable mutations (Liang et al, 2019;Chen and Arnold, 2020;Jeong et al, 2020;Markel et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Light-Driven CO2 Reduction by Co-Cytochrome b562

Alcala-Torano

Halloran

Gwerder

et al. 2021

Front. Mol. Biosci.

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The current trend in atmospheric carbon dioxide concentrations is causing increasing concerns for its environmental impacts, and spurring the developments of sustainable methods to reduce CO2 to usable molecules. We report the light-driven CO2 reduction in water in mild conditions by artificial protein catalysts based on cytochrome b562 and incorporating cobalt protoporphyrin IX as cofactor. Incorporation into the protein scaffolds enhances the intrinsic reactivity of the cobalt porphyrin toward proton reduction and CO generation. Mutations around the binding site modulate the activity of the enzyme, pointing to the possibility of further improving catalytic activity through rational design or directed evolution.

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Proteins as diverse, efficient, and evolvable scaffolds for artificial metalloenzymes

Abstract: We have extracted and categorized the desirable properties of proteins that are adapted as the scaffolds for artificial metalloenzymes.

Cited by 33 publications

References 150 publications

Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

Protein Modifications: From Chemoselective Probes to Novel Biocatalysts

Light-Driven CO2 Reduction by Co-Cytochrome b562

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