Intracellular Unnatural Catalysis Enabled by an Artificial Metalloenzyme

Methods Mol Biol. 2021:2312:287-300. doi: 10.1007/978-1-0716-1441-9_17.

Abstract

Artificial metalloenzymes, constructed by incorporating a synthetic catalyst into the internal spaces of a protein scaffold, can perform noncanonical chemical transformations that are not possible using natural enzymes. The addition of cell-permeable modules to artificial metalloenzymes allows for noncanonical catalysis to be implemented as a function of mammalian cells. In this chapter, we describe a protocol for controlling cellular function through a cascade consisting of an artificial metalloenzyme and a gene-circuit engineered via synthetic biology.

Keywords: Allylic dealkylation; Artificial metalloenzyme; Cell-penetrating polydisulfides; Gene switch; Intracellular catalysis; Ruthenium complex; Streptavidin; Thyroid hormone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / chemistry
  • Catalysis
  • Cell Culture Techniques
  • Cell Engineering*
  • Enzymes / genetics
  • Enzymes / metabolism*
  • Gene Expression Regulation, Enzymologic
  • HEK293 Cells
  • Humans
  • Metalloproteins / genetics
  • Metalloproteins / metabolism*
  • Protein Engineering*
  • Streptavidin / chemistry
  • Substrate Specificity
  • Synthetic Biology*
  • Transfection

Substances

  • Enzymes
  • Metalloproteins
  • Biotin
  • Streptavidin