Light-driven post-translational installation of reactive protein side chains

Nature. 2020 Sep;585(7826):530-537. doi: 10.1038/s41586-020-2733-7. Epub 2020 Sep 23.

Abstract

Post-translational modifications (PTMs) greatly expand the structures and functions of proteins in nature1,2. Although synthetic protein functionalization strategies allow mimicry of PTMs3,4, as well as formation of unnatural protein variants with diverse potential functions, including drug carrying5, tracking, imaging6 and partner crosslinking7, the range of functional groups that can be introduced remains limited. Here we describe the visible-light-driven installation of side chains at dehydroalanine residues in proteins through the formation of carbon-centred radicals that allow C-C bond formation in water. Control of the reaction redox allows site-selective modification with good conversions and reduced protein damage. In situ generation of boronic acid catechol ester derivatives generates RH2C radicals that form the native (β-CH2-γ-CH2) linkage of natural residues and PTMs, whereas in situ potentiation of pyridylsulfonyl derivatives by Fe(II) generates RF2C radicals that form equivalent β-CH2-γ-CF2 linkages bearing difluoromethylene labels. These reactions are chemically tolerant and incorporate a wide range of functionalities (more than 50 unique residues/side chains) into diverse protein scaffolds and sites. Initiation can be applied chemoselectively in the presence of sensitive groups in the radical precursors, enabling installation of previously incompatible side chains. The resulting protein function and reactivity are used to install radical precursors for homolytic on-protein radical generation; to study enzyme function with natural, unnatural and CF2-labelled post-translationally modified protein substrates via simultaneous sensing of both chemo- and stereoselectivity; and to create generalized 'alkylator proteins' with a spectrum of heterolytic covalent-bond-forming activity (that is, reacting diversely with small molecules at one extreme or selectively with protein targets through good mimicry at the other). Post-translational access to such reactions and chemical groups on proteins could be useful in both revealing and creating protein function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / analogs & derivatives
  • Alanine / chemistry
  • Alanine / metabolism
  • Binding Sites
  • Carbon / chemistry
  • Carbon / metabolism
  • Enzymes / chemistry
  • Enzymes / metabolism
  • Esters / chemical synthesis
  • Esters / chemistry
  • HeLa Cells
  • Humans
  • Hydrocarbons, Fluorinated / chemistry
  • Hydrocarbons, Fluorinated / metabolism
  • Indicators and Reagents / chemistry
  • Light*
  • Oxidation-Reduction
  • Photochemical Processes / radiation effects
  • Protein Interaction Domains and Motifs
  • Protein Processing, Post-Translational / radiation effects*
  • Proteins / chemistry*
  • Proteins / metabolism*

Substances

  • Enzymes
  • Esters
  • Hydrocarbons, Fluorinated
  • Indicators and Reagents
  • Proteins
  • Carbon
  • difluoromethane
  • dehydroalanine
  • Alanine