Design and synthesis of heteronuclear metalloenzymes involved in multi-electron and multi-proton processes

Many enzymes which catalyze reactions essential to sustainable energy, human health, and the environment also contain heteronuclear metal centers and involve multi-electron and multi-proton processes. These processes include respiration, biomass conversion, use of biofuels, and the global nitrogen/carbon/sulfur cycles. Despite their importance, most of these enzymes are not well understood in comparison with mono- or homonuclear metalloenzymes, in part due to the complexity inherent in the presence of different metal ions in close proximity.

To address this issue, we have developed a biosynthetic approach to synthesize structural and functional mimics of cytochrome c oxidase, nitric oxide reductase, sulfite reductase and manganese peroxidase. By tuning the secondary sphere interactions, we have made protein models with activity comparable to that of native enzymes. Using these small and robust protein models, we have obtained deeper insight into structural features responsible for high activity.

  1. Ambika Bhagi-Damodaran, Igor Petrik, Yi Lu, Using Biosynthetic Models of Heme-Copper Oxidase and Nitric Oxide Reductase in Myoglobin to Elucidate Structural Features Responsible for Enzymatic Activities. Isr. J. Chem. 56, 773-790 (2016).
  2. Yang Yu, Chang Cui, Xiaohong Liu, Igor Petrik, Jiangyun Wang, Yi Lu, “A Designed Metalloenzyme Achieving the Catalytic Rate of a Native Enzyme” J. Am. Chem. Soc. 137, (36) 11570-11573 (2015).
  3. Ambika Bhagi-Damodaran, Matthew A. Michael, Qianhong Zhu, Julian Reed, Braddock A. Sandoval, Evan N. Mirts, Saumen Chakraborty, Pierre Moenne-Loccoz, Yong Zhang, Yi Lu, “Why copper is preferred over iron for oxygen activation and reduction in heme-copper oxidases” Nature Chemistry 9, 257-263 (2017).
  4. Natasha Yeung, Ying-Wu Lin, Yi-Gui Gao, Xuan Zhao, Brandy S. Russell, Lanyu Lei, Kyle D. Miner, Howard Robinson, Yi Lu, “Rational design of a structural and functional nitric oxide reductase” Nature 462, 1079-1082 (2009).
  5. Ying-Wu Lin, Natasha Yeung, Yi-Gui Gao, Kyle D. Miner, Shiliang Tian, Howard Robinson, Yi Lu, “Roles of glutamates and metal ions in a rationally designed nitric oxide reductase based on myoglobin”, Proceedings of the National Academy of Sciences 107, 8581-8586 (2010).
  6. Parisa Hosseinzadeh, Evan N. Mirts, Thomas D. Pfister, Yi-Gui Gao, Christopher Mayne, Howard Robinson, Emad Tajkhorshid, Yi Lu, “Enhancing Mn(II)-binding and manganese peroxidase activity in a designed cytochrome c peroxidase through fine-tuning secondary sphere interactions” Biochemistry 55, (10) 1494-1502 (2016).