Abstract
It has been suggested that in mammalian cells histidine residues in proteins may become as frequently phosphorylated as serine, threonine and tyrosine, and may play a key role in mammalian signaling. Here we applied a robust workflow that earlier allowed us to detect histidine phosphorylation in bacteria unambiguously, to probe for histidine phosphorylation in four human cell lines. Initially, seemingly hundreds of protein histidine phosphorylations were picked up in all studied human cell lines. However, careful examination of the data, and several control experiments, led us to the conclusion that >99% of these initially assigned pHis sites were not genuine, and should be site localized to neighboring Ser/Thr residues. Nevertheless, our methods are selective enough to detect just a handful of genuine pHis sites in mammalian cells, representing well-known enzymatic intermediates. Consequently, we do not find any evidence in our data supporting that protein histidine phosphorylation plays a role in mammalian signaling.
| Original language | English |
|---|---|
| Pages (from-to) | 827-828 |
| Journal | Nature Methods |
| Volume | 19 |
| Issue number | 7 |
| Early online date | 20 Jun 2022 |
| DOIs | |
| Publication status | Published - Jul 2022 |
Bibliographical note
Funding Information:We thank B. van Breukelen for assistance with preparing the figures. This research received funding through the Netherlands Organization for Scientific Research through a VIDI grant (project no. 723.013.008) to S.L. and the Spinoza Award SPI.2017.028 to A.J.R.H. This project received additional funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 686547 (EPIC-XS).
Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.