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Native chemical ligation at methionine bioisostere norleucine allows for N-terminal chemical protein ligation.

Bo-Tao XinBianca D M van TolHuib OvaaPaul P Geurink
Published in: Organic & biomolecular chemistry (2019)
The development of γ-thionorleucine (ThioNle) as a handle for native chemical ligation-desulfurization is reported here. ThioNle is a new addition to the expanding thiolated amino acid toolbox and serves as a methionine substitute in NCL with the advantage that it lacks the undesirable oxidation-prone thioether moiety. Its usefulness for N-terminal ubiquitination is demonstrated by efficient preparation of fully synthetic linear diubiquitin with preserved protein folding compared to the expressed material. Interestingly, gel-based deubiquitinating assays revealed that the methionine to norleucine substitution did affect diubiquitin cleavage, which may indicate a more profound role for methionine in the interaction between ubiquitin and the deubiquitinating enzymes than has been known so far.
Keyphrases
  • amino acid
  • small molecule
  • high throughput
  • molecular dynamics simulations
  • nitric oxide
  • transcription factor
  • mass spectrometry
  • binding protein
  • molecularly imprinted