A Designed Functional Metalloenzyme that Reduces O<sub>2</sub> to H<sub>2</sub>O with Over One Thousand Turnovers

Miner, Kyle D.; Mukherjee, Arnab; Gao, Yu; Null, Eric L.; Petrík, Igor; Zhao, Xuan; Yeung, Natasha; Robinson, Howard; Lu, Yi

doi:10.1002/ange.201201981

Cited by 19 publications

(13 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To address this problem, they introduced Tyr residues at two different positions (33 and 65) close to the histidines of the Cu B site (Figure 6). The resulting enzymes then efficiently reduced O 2 to water (Gly65Tyr Cu B Mb with over 1000 turnovers) with minimal release of superoxide or peroxide 41…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Rational Heme Protein Design: All Roads Lead to Rome

Lin

Sawyer

Wang

2013

Chemistry An Asian Journal

View full text Add to dashboard Cite

Heme proteins are among the most abundant and important metalloproteins, exerting diverse biological functions including oxygen transport, small molecule sensing, selective CH bond activation, nitrite reduction, and electron transfer. Rational heme protein designs focus on the modification of the heme‐binding active site and the heme group, protein hybridization and domain swapping, and de novo design. These strategies not only provide us with unique advantages for illustrating the structure–property–reactivity–function (SPRF) relationship of heme proteins in nature but also endow us with the ability to create novel biocatalysts and biosensors.

show abstract

Section: Introductionmentioning

confidence: 99%

“… (A) Crystal structure of F33Y Cu B Mb with Cu II bound (PDB entry 4FWY41). (B) A modeling structure of G65Y Cu B Mb [41] .…”

Section: Introductionmentioning

confidence: 99%

Rational Heme Protein Design: All Roads Lead to Rome

Lin

Sawyer

Wang

2013

Chemistry An Asian Journal

View full text Add to dashboard Cite

show abstract

“…In this reaction, the presence of a tyrosine residue that is covalently linked to one of the histidine ligands to the Cu center appears to play a role in electron and proton transfers in HCO 22. In the next protein design step, a tyrosine residue was introduced at the active site of Cu⋅L29H/F43HMb to model the role of tyrosine in the reduction of O 2 by HCO 23. The introduction of Tyr at position 65 improved the reaction.…”

Section: Small Proteins For Fine‐tuning Of the Active Sitementioning

confidence: 99%

Artificial Metalloenzymes Constructed From Hierarchically‐Assembled Proteins

Ueno

Tabe

Tanaka

2013

Chemistry An Asian Journal

View full text Add to dashboard Cite

The design of artificial metalloenzymes has become an important topic in biological chemistry and inorganic chemistry due to the potential applications of artificial metalloenzymes in nanoscience and biotechnology. One of the general methods used to produce artificially metalloenzymes involves the encapsulation of non-natural metal cofactors within protein scaffolds. This method has been used in the construction of small artificial metalloproteins with high activity and selectivity. However, the important roles of protein assemblies have not yet been systematically investigated in this field, even though natural enzymatic systems employ protein assemblies as molecular scaffolds for elaborate enzymatic reactions. In recent years, the above-mentioned general strategy has been applied to functionalize protein assemblies such as protein cages and protein crystals. These assembled structures form confined interior environments, which can be used to accommodate metal complex catalysts and to prepare metal nanoparticles. The development of artificial metalloenzymes with hierarchically-assembled proteins would enable us to provide powerful tools for industrial and biological applications. In this Focus Review, we discuss the most significant recent research in this field as well as future directions.

show abstract

“…[12] We then introduced atyrosine through F33Y mutation next to histidine ligands to model the conserved tyrosine in HCO. [13] Ther esultant mutant named F33Y-Cu B Mb ( Figure 1A) mimicked HCOs functionally,a si tc ould selectively reduce oxygen to water with hundreds of turnovers. [14] We further modulated heme E8 8' of F33Y-Cu B Mb by 210 mV ( Figure 1B) via tuning of hydrogen bonding to heme iron (through S92A mutation) and using non-native heme cofactors with increased E8 8',s uch as monoformyl (MF-) and diformyl (DF-) hemes.…”

mentioning

confidence: 99%

Insights Into How Heme Reduction Potentials Modulate Enzymatic Activities of a Myoglobin‐based Functional Oxidase

et al. 2017

Self Cite

View full text Add to dashboard Cite

Heme-copper oxidase (HCO) is a class of respiratory enzymes that use a heme-copper center to catalyze O2 reduction to H2O. While heme reduction potential (E°′) of different HCO types has been found to vary >500 mV, its impact on HCO activity remains poorly understood. Here, we use a set of myoglobin-based functional HCO models to investigate the mechanism by which heme E°′ modulates oxidase activity. Rapid stopped-flow kinetic measurements show that increasing heme E°′ by ca. 210 mV results in increases in electron transfer (ET) rates by 30-fold, rate of O2 binding by 12-fold, O2 dissociation by 35-fold, while decreasing O2 affinity by 3-fold. Theoretical calculations reveal that E°′ modulation has significant implications on electronic charge of both heme iron and O2, resulting in increased O2 dissociation and reduced O2 affinity at high E°′ values. This work suggests fine-tuning E°′ in HCOs and other heme enzymes can modulate their substrate affinity, ET rate and enzymatic activity.

show abstract

A Designed Functional Metalloenzyme that Reduces O₂ to H₂O with Over One Thousand Turnovers

Cited by 19 publications

References 44 publications

Rational Heme Protein Design: All Roads Lead to Rome

Rational Heme Protein Design: All Roads Lead to Rome

Artificial Metalloenzymes Constructed From Hierarchically‐Assembled Proteins

Insights Into How Heme Reduction Potentials Modulate Enzymatic Activities of a Myoglobin‐based Functional Oxidase

Contact Info

Product

Resources

About

A Designed Functional Metalloenzyme that Reduces O2 to H2O with Over One Thousand Turnovers

Cited by 19 publications

References 44 publications

Rational Heme Protein Design: All Roads Lead to Rome

Rational Heme Protein Design: All Roads Lead to Rome

Artificial Metalloenzymes Constructed From Hierarchically‐Assembled Proteins

Insights Into How Heme Reduction Potentials Modulate Enzymatic Activities of a Myoglobin‐based Functional Oxidase

Contact Info

Product

Resources

About

A Designed Functional Metalloenzyme that Reduces O₂ to H₂O with Over One Thousand Turnovers