Threonine eliminylation by bacterial phosphothreonine lyases rapidly causes cross-linking of mitogen-activated protein kinase (MAPK) in live cells
| Autor: | Ida Chiara Guerrera, Benoit M. Meijer, Laurence Arbibe, Philippe J. Sansonetti, Françoise Baleux, Cerina Chhuon, Joanna Lipecka, Caroline Reisacher, Suk Min Jang, Christian Muchardt |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
MAPK/ERK pathway Proteomics Threonine Genomics and Proteomics MAP Kinase Signaling System Carbon-Oxygen Lyases Plasma protein binding Protein aggregation Biochemistry p38 Mitogen-Activated Protein Kinases Antibodies Cell Line Substrate Specificity 03 medical and health sciences Mice 0302 clinical medicine Bacterial Proteins Type III Secretion Systems Animals Humans Secretion Extracellular Signal-Regulated MAP Kinases Molecular Biology chemistry.chemical_classification Alanine biology Aminobutyrates Cell Biology Amino acid 030104 developmental biology chemistry Mitogen-activated protein kinase biology.protein Phosphothreonine Shigella Caco-2 Cells 030217 neurology & neurosurgery HeLa Cells Protein Binding |
| Zdroj: | The Journal of biological chemistry. 292(19) |
| ISSN: | 1083-351X |
| Popis: | Old long-lived proteins contain dehydroalanine (Dha) and dehydrobutyrine (Dhb), two amino acids engendered by dehydration of serines and threonines, respectively. Although these residues have a suspected role in protein cross-linking and aggregation, their direct implication has yet to be determined. Here, we have taken advantage of the ability of the enteropathogen Shigella to convert the phosphothreonine residue of the pT-X-pY consensus sequence of ERK and p38 into Dhb and followed the impact of dehydration on the fate of the two MAPKs. To that end, we have generated the first antibodies recognizing Dhb-modified proteins and allowing tracing them as they form. We showed that Dhb modifications accumulate in a long-lasting manner in Shigella-infected cells, causing subsequent formation of covalent cross-links of MAPKs. Moreover, the Dhb signal correlates precisely with the activation of the Shigella type III secretion apparatus, thus evidencing injectisome activity. This observation is the first to document a causal link between Dhb formation and protein cross-linking in live cells. Detection of eliminylation is a new avenue to phosphoproteome regulation in eukaryotes that will be instrumental for the development of type III secretion inhibitors. |
| Databáze: | OpenAIRE |
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