Introducing a 2-His-1-Glu Nonheme Iron Center into Myoglobin Confers Nitric Oxide Reductase Activity

Lin, Ying Wu; Yeung, Natasha; Gao, Yi; Miner, Kyle D.; Liu, Lei; Robinson, Howard; Lu, Yi

doi:10.1021/ja103516n

Cited by 59 publications

(50 citation statements)

References 23 publications

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“…Making a closer observation of the cNOR non-heme Fe B site, which presents a three histidine and a glutamate coordination, it does not seem probable the formation of a Fe B -dinitrosyl complex, due to the high number of coordinating ligands. Also, studies on the modified sperm whale myoglobin indicated that only two histidines and a glutamate residue are essential for the non-heme Fe B coordination and consequent NO reduction [57].…”

Section: New Insights On the Catalytic Mechanism Of No Reductionmentioning

confidence: 99%

Steady-state kinetics with nitric oxide reductase (NOR): New considerations on substrate inhibition profile and catalytic mechanism

Duarte

Cordas

Moura

et al. 2014

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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Nitric oxide reductase (NOR) from denitrifying bacteria is an integral membrane protein that catalyses the two electron reduction of NO to N2O, as part of the denitrification process, being responsible for an exclusive reaction, the NN bond formation, the key step of this metabolic pathway. Additionally, this class of enzymes also presents residual oxidoreductase activity, reducing O2 to H2O in a four electron/proton reaction. In this work we report, for the first time, steady-state kinetics with the Pseudomonas nautica NOR, either in the presence of its physiological electron donor (cyt. c552) or immobilised on a graphite electrode surface, in the presence of its known substrates, namely NO or O2. The obtained results show that the enzyme has high affinity for its natural substrate, NO, and different kinetic profiles according to the electron donor used. The kinetic data, as shown by the pH dependence, is modelled by ionisable amino acid residues nearby the di-nuclear catalytic site. The catalytic mechanism is revised and a mononitrosyl-non-heme Fe complex (FeB(II)-NO) species is favoured as the first catalytic intermediate involved on the NO reduction.

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Section: New Insights On the Catalytic Mechanism Of No Reductionmentioning

confidence: 99%

Steady-state kinetics with nitric oxide reductase (NOR): New considerations on substrate inhibition profile and catalytic mechanism

Duarte

Cordas

Moura

et al. 2014

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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“…During the last few decades, swMb has been well studied as a represent heme protein. In addition, it has been widely used as a protein model for rational design of artificial heme proteins with novel functions [1,4,[8][9][10][11]. Comparatively, the structureactivity relationship has not been fully understood for the unique apMb [6,[12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Dynamics comparison of two myoglobins with a distinct heme active site

Lin

Liao

2011

J Mol Model

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Sperm whale myoglobin (swMb) is a well-studied heme protein, both experimentally and theoretically. Comparatively, little attention has been paid to another member of Mb family, Aplysia limacina myoglobin (apMb). swMb and apMb have the same overall structure and perform the same biological function, i.e., O(2) carrier, while using a distinct heme active site. To provide insights into the structure-function relationship for these two Mbs, we herein made a comparison in terms of their dynamics properties using molecular dynamics simulation. We analyzed the overall structure and protein motions, as well as the intramolecular contacts, namely salt-bridges and hydrogen bonds, especially the interactions between the heme propionate groups and the polypeptide chain. The internal cavities in apMb were also compared to the well-known xenon and other cavities in swMb. Based on current simulations, we propose a unique "arginine gate" for apMb, which has a similar function to the histidine gate observed for swMb in previous studies. This study provides valuable information for understanding the homology of heme proteins, and also aids in rational design of structural and functional heme proteins by alternating the heme active site.

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“…General purification of our designed heteronuclear metalloproteins will not be discussed in great detail here, but generally falls into two categories: inclusion body and cytosolic purification. The inclusion body protocol is used extensively in models such as Cu B Mb and Fe B Mb and has been covered previously in those works (Springer & Sligar, 1987, Sigman et al, 2000, Sigman et al, 2000, Sigman et al, 2003, Pfister et al, 2005, Zhao et al, 2005, Zhao et al, 2006, Yeung et al, 2009, Lin et al, 2010, Lin et al, 2010, Miner et al, 2012, Chakraborty et al, 2014, Bhagi-Damodaran et al, 2014, Petrik et al, 2016). Additionally, cytosolic purification of C c P and its rationally designed mutants has also been described in great detail by our group (Yeung et al, 1997, Sigman et al, 1999, Feng et al, 2003, Pfister et al, 2007, Miner et al, 2014, Hosseinzadeh et al, 2016).…”

Section: Step 2: Purification and Structural Characterization Of Ratimentioning

confidence: 99%

Design of Heteronuclear Metalloenzymes

Bhagi‐Damodaran

Hosseinzadeh

Mirts

et al. 2016

Methods in Enzymology

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Heteronuclear metalloenzymes catalyze some of the most fundamentally interesting and practically useful reactions in nature. However, the presence of two or more metal ions in close proximity in these enzymes makes them more difficult to prepare and study than homonuclear metalloenzymes. To meet these challenges, heteronuclear metal centers have been designed into small and stable proteins with rigid scaffolds to understand how these heteronuclear centers are constructed and the mechanism of their function. This chapter describes methods for designing heterobinuclear metal centers in a protein scaffold by giving specific examples of a few heme-nonheme bimetallic centers engineered in myoglobin and cytochrome c peroxidase. We provide step-by-step procedure on how to choose the protein scaffold, design a heterobinuclear metal center in the protein computationally, incorporate metal centers in the protein and characterize the resulting metalloprotein, both structurally and functionally. Finally, we discuss how an initial design can be further improved by rationally tuning its secondary coordination sphere, electron/proton transfer rates, and the substrate affinity.

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Introducing a 2-His-1-Glu Nonheme Iron Center into Myoglobin Confers Nitric Oxide Reductase Activity

Cited by 59 publications

References 23 publications

Steady-state kinetics with nitric oxide reductase (NOR): New considerations on substrate inhibition profile and catalytic mechanism

Steady-state kinetics with nitric oxide reductase (NOR): New considerations on substrate inhibition profile and catalytic mechanism

Dynamics comparison of two myoglobins with a distinct heme active site

Design of Heteronuclear Metalloenzymes

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