Design of Heteronuclear Metalloenzymes

A. Bhagi-Damodaran, P. Hosseinzadeh, E. Mirts, J. Reed, I. D. Petrik, Y. Lu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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 procedures on how to choose the protein scaffold, design a heterobinuclear metal center in the protein scaffold computationally, incorporate metal ions into the protein, and characterize the resulting metalloproteins, 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.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages501-537
Number of pages37
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Enzymology
Volume580
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Biomimetic models
  • Biosynthetic models
  • Heme–copper oxidase
  • Manganese peroxidase
  • Metalloprotein design
  • Nitric oxide reductase
  • Oxygen activation
  • Protein design
  • Protein engineering
  • Secondary sphere interactions

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Design of Heteronuclear Metalloenzymes'. Together they form a unique fingerprint.

  • Cite this

    Bhagi-Damodaran, A., Hosseinzadeh, P., Mirts, E., Reed, J., Petrik, I. D., & Lu, Y. (2016). Design of Heteronuclear Metalloenzymes. In Methods in Enzymology (pp. 501-537). (Methods in Enzymology; Vol. 580). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.05.050