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Substrate recognition by the Pseudomonas aeruginosa EF-Tu–modifying methyltransferase EftM
| Content Provider | Scilit |
|---|---|
| Author | Kuiper, Emily G. Dey, Debayan LaMore, Paige A. Owings, Joshua P. Prezioso, Samantha M. Goldberg, Joanna B. Conn, Graeme L. |
| Copyright Year | 2019 |
| Description | Journal: Journal of Biological Chemistry The opportunistic bacterial pathogen Pseudomonas aeruginosa is a leading cause of serious infections in individuals with cystic fibrosis, compromised immune systems, or severe burns. P. aeruginosa adhesion to host epithelial cells is enhanced by surface-exposed translation elongation factor EF-Tu carrying a Lys-5 trimethylation, incorporated by the methyltransferase EftM. Thus, the EF-Tu modification by EftM may represent a target to prevent P. aeruginosa infections in vulnerable individuals. Here, we extend our understanding of EftM activity by defining the molecular mechanism by which it recognizes EF-Tu. Acting on the observation that EftM can bind to EF-Tu lacking its N-terminal peptide (encompassing the Lys-5 target site), we generated an EftM homology model and used it in protein/protein docking studies to predict EftM/EF-Tu interactions. Using site-directed mutagenesis of residues in both proteins, coupled with binding and methyltransferase activity assays, we experimentally validated the predicted protein/protein interface. We also show that EftM cannot methylate the isolated N-terminal EF-Tu peptide and that binding-induced conformational changes in EftM are likely needed to enable placement of the first 5–6 amino acids of EF-Tu into a conserved peptide-binding channel in EftM. In this channel, a group of residues that are highly conserved in EftM proteins position the N-terminal sequence to facilitate Lys-5 modification. Our findings reveal that EftM employs molecular strategies for substrate recognition common among both class I (Rossmann fold) and class II (SET domain) methyltransferases and pave the way for studies seeking a deeper understanding of EftM's mechanism of action on EF-Tu. |
| Related Links | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6937583/pdf http://www.jbc.org/content/294/52/20109.full.pdf |
| Ending Page | 20121 |
| Page Count | 13 |
| Starting Page | 20109 |
| ISSN | 00219258 |
| e-ISSN | 1083351X |
| DOI | 10.1074/jbc.ra119.011213 |
| Journal | Journal of Biological Chemistry |
| Issue Number | 52 |
| Volume Number | 294 |
| Language | English |
| Publisher | Elsevier BV |
| Publisher Date | 2019-12-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Biological Chemistry Biochemistry and Molecular Biology Pseudomonas Aeruginosa (p. Aeruginosa) S-adenosylmethionine (sam) Post-translational Modification (ptm) Protein Methylation Substrate Specificity Translation Elongation Factor |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
