Probing Backbone Hydrogen Bonds in Proteins by Amide-to-Ester Mutations

Chembiochem. 2018 Oct 18;19(20):2136-2145. doi: 10.1002/cbic.201800350. Epub 2018 Oct 1.


All proteins contain characteristic backbones formed of consecutive amide bonds, which can engage in hydrogen bonds. However, the importance of these is not easily addressed by conventional technologies that only allow for side-chain substitutions. By contrast, technologies such as nonsense suppression mutagenesis and protein ligation allow for manipulation of the protein backbone. In particular, replacing the backbone amide groups with ester groups, that is, amide-to-ester mutations, is a powerful tool to examine backbone-mediated hydrogen bonds. In this minireview, we showcase examples of how amide-to-ester mutations can be used to uncover pivotal roles of backbone-mediated hydrogen bonds in protein recognition, folding, function, and structure.

Keywords: amide-to-ester mutations; hydrogen bonds; protein backbone; proteins; structure and function.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Amides / chemistry*
  • Codon, Nonsense*
  • Esters / chemistry*
  • Hydrogen Bonding
  • Mutagenesis
  • Protein Conformation
  • Protein Folding
  • Proteins / chemistry*
  • Proteins / genetics*


  • Amides
  • Codon, Nonsense
  • Esters
  • Proteins