Reference - Detail
| RRC ID | 71365 |
|---|---|
| Author | Liu A, Si Y, Dong SH, Mahanta N, Penkala HN, Nair SK, Mitchell DA. |
| Title | Functional elucidation of TfuA in peptide backbone thioamidation. |
| Journal | Nat Chem Biol |
| Abstract |
YcaO enzymes catalyze several post-translational modifications on peptide substrates, including thioamidation, which substitutes an amide oxygen with sulfur. Most predicted thioamide-forming YcaO enzymes are encoded adjacent to TfuA, which when present, is required for thioamidation. While activation of the peptide amide backbone is well established for YcaO enzymes, the function of TfuA has remained enigmatic. Here we characterize the TfuA protein involved in methyl-coenzyme M reductase thioamidation and demonstrate that TfuA catalyzes the hydrolysis of thiocarboxylated ThiS (ThiS-COSH), a proteinaceous sulfur donor, and enhances the affinity of YcaO toward the thioamidation substrate. We also report a crystal structure of a TfuA, which displays a new protein fold. Our structural and mutational analyses of TfuA have uncovered conserved binding interfaces with YcaO and ThiS in addition to revealing a hydrolase-like active site featuring a Ser-Lys catalytic pair. |
| Volume | 17(5) |
| Pages | 585-592 |
| Published | 2021-5-1 |
| DOI | 10.1038/s41589-021-00771-0 |
| PII | 10.1038/s41589-021-00771-0 |
| PMID | 33707784 |
| PMC | PMC8084935 |
| MeSH | Archaeal Proteins / chemistry* Archaeal Proteins / genetics Archaeal Proteins / metabolism Binding Sites Cloning, Molecular Crystallography, X-Ray Escherichia coli / genetics Escherichia coli / metabolism Euryarchaeota / enzymology* Euryarchaeota / genetics Gene Expression Genetic Vectors / chemistry Genetic Vectors / metabolism Histidine / chemistry Histidine / genetics Histidine / metabolism Kinetics Mannose-Binding Lectin / chemistry Mannose-Binding Lectin / genetics Mannose-Binding Lectin / metabolism Methanobacteriaceae / enzymology* Methanobacteriaceae / genetics Methanocaldococcus / enzymology* Methanocaldococcus / genetics Models, Molecular Mutation Oligopeptides / chemistry Oligopeptides / genetics Oligopeptides / metabolism Oxidoreductases / chemistry* Oxidoreductases / genetics Oxidoreductases / metabolism Peptides / chemistry Peptides / genetics Peptides / metabolism Protein Binding Protein Conformation, alpha-Helical Protein Conformation, beta-Strand Protein Interaction Domains and Motifs Protein Processing, Post-Translational Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Substrate Specificity Thioamides / chemistry* Thioamides / metabolism |
| Resource | |
| General Microbes | JCM14679 |