1. A Chemical Probe for Dehydrobutyrine.
- Author
-
Chambers KA, Abularrage NS, Hill CJ, Khan IH, and Scheck RA
- Subjects
- Alanine analogs & derivatives, Alanine chemistry, Amines chemistry, Bacteria enzymology, Bacterial Infections enzymology, Biomarkers, Histones chemistry, Humans, Lyases chemistry, Phosphines, Phosphothreonine, Protein Processing, Post-Translational, Sulfhydryl Compounds chemistry, Aminobutyrates chemistry
- Abstract
Bacterial phosphothreonine lyases, or phospholyases, catalyze a unique post-translational modification that introduces dehydrobutyrine (Dhb) or dehydroalanine (Dha) in place of phosphothreonine or phosphoserine residues, respectively. We report the use of a phospha-Michael reaction to label proteins and peptides modified with Dha or Dhb. We demonstrate that a nucleophilic phosphine probe is able to modify Dhb-containing proteins and peptides that were recalcitrant to reaction with thiol or amine nucleophiles under mild aqueous conditions. Furthermore, we used this reaction to detect multiple Dhb-modified proteins in mammalian cell lysates, including histone H3, a previously unknown target of phospholyases. This method should prove useful for identifying new phospholyase targets, profiling the biomarkers of bacterial infection, and developing enzyme-mediated strategies for bioorthogonal labeling in living cells., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2020
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