Heme redox potentials hold the key to reactivity differences between nitric oxide reductase and heme-copper oxidase

Ambika Bhagi-Damodaran, Julian H. Reed, Qianhong Zhu, Yelu Shi, Parisa Hosseinzadeh, Braddock A. Sandoval, Kevin A. Harnden, Shuyan Wang, Madeline R. Sponholtz, Evan N. Mirts, Sudharsan Dwaraknath, Yong Zhang, Pierre Moenne-Loccoz, Yi Lu

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Despite high structural homology between NO reductases (NORs) and heme-copper oxidases (HCOs), factors governing their reaction specificity remain to be understood. Using a myoglobinbased model of NOR (FeBMb) and tuning its heme redox potentials (E°′) to cover the native NOR range, through manipulating hydrogen bonding to the proximal histidine ligand and replacing heme b with monoformyl (MF-) or diformyl (DF-) hemes, we herein demonstrate that the E°′ holds the key to reactivity differences between NOR and HCO. Detailed electrochemical, kinetic, and vibrational spectroscopic studies, in tandem with density functional theory calculations, demonstrate a strong influence of heme E°′ on NO reduction. Decreasing E°′ from +148 to -130 mV significantly impacts electronic properties of the NOR mimics, resulting in 180- and 633-fold enhancements in NO association and hemenitrosyl decay rates, respectively. Our results indicate that NORs exhibit finely tuned E°′ that maximizes their enzymatic efficiency and helps achieve a balance between opposite factors: fast NO binding and decay of dinitrosyl species facilitated by low E°′ and fast electron transfer facilitated by high E°′. Only when E°′ is optimally tuned in FeBMb(MF-heme) for NO binding, heme-nitrosyl decay, and electron transfer does the protein achieve multiple (>35) turnovers, previously not achieved by synthetic or enzymebased NOR models. This also explains a long-standing question in bioenergetics of selective cross-reactivity in HCOs. Only HCOs with heme E°′ in a similar range as NORs (between -59 and 200 mV) exhibit NOR reactivity. Thus, our work demonstrates efficient tuning of E°′ in various metalloproteins for their optimal functionality.

Original languageEnglish (US)
Pages (from-to)6195-6200
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number24
DOIs
StatePublished - Jun 12 2018

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Heme
Oxidation-Reduction
Oxidoreductases
nitric-oxide reductase
copper oxidase
Metalloproteins
Electrons
Hydrogen Bonding
Histidine
Energy Metabolism
Ligands

Keywords

  • Biomimetics
  • Heme-copper oxidase
  • Metalloprotein design
  • Nitric oxide reductase
  • Redox potentials

ASJC Scopus subject areas

  • General

Cite this

Heme redox potentials hold the key to reactivity differences between nitric oxide reductase and heme-copper oxidase. / Bhagi-Damodaran, Ambika; Reed, Julian H.; Zhu, Qianhong; Shi, Yelu; Hosseinzadeh, Parisa; Sandoval, Braddock A.; Harnden, Kevin A.; Wang, Shuyan; Sponholtz, Madeline R.; Mirts, Evan N.; Dwaraknath, Sudharsan; Zhang, Yong; Moenne-Loccoz, Pierre; Lu, Yi.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 24, 12.06.2018, p. 6195-6200.

Research output: Contribution to journalArticle

Bhagi-Damodaran, A, Reed, JH, Zhu, Q, Shi, Y, Hosseinzadeh, P, Sandoval, BA, Harnden, KA, Wang, S, Sponholtz, MR, Mirts, EN, Dwaraknath, S, Zhang, Y, Moenne-Loccoz, P & Lu, Y 2018, 'Heme redox potentials hold the key to reactivity differences between nitric oxide reductase and heme-copper oxidase', Proceedings of the National Academy of Sciences of the United States of America, vol. 115, no. 24, pp. 6195-6200. https://doi.org/10.1073/pnas.1720298115
Bhagi-Damodaran, Ambika ; Reed, Julian H. ; Zhu, Qianhong ; Shi, Yelu ; Hosseinzadeh, Parisa ; Sandoval, Braddock A. ; Harnden, Kevin A. ; Wang, Shuyan ; Sponholtz, Madeline R. ; Mirts, Evan N. ; Dwaraknath, Sudharsan ; Zhang, Yong ; Moenne-Loccoz, Pierre ; Lu, Yi. / Heme redox potentials hold the key to reactivity differences between nitric oxide reductase and heme-copper oxidase. In: Proceedings of the National Academy of Sciences of the United States of America. 2018 ; Vol. 115, No. 24. pp. 6195-6200.
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AU - Reed, Julian H.

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AU - Shi, Yelu

AU - Hosseinzadeh, Parisa

AU - Sandoval, Braddock A.

AU - Harnden, Kevin A.

AU - Wang, Shuyan

AU - Sponholtz, Madeline R.

AU - Mirts, Evan N.

AU - Dwaraknath, Sudharsan

AU - Zhang, Yong

AU - Moenne-Loccoz, Pierre

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