Sequence–structure–function analysis of the bifunctional enzyme MnmC that catalyses the last two steps in the biosynthesis of hypermodified nucleoside mnm5s2U in tRNA
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| Titel: | Sequence–structure–function analysis of the bifunctional enzyme MnmC that catalyses the last two steps in the biosynthesis of hypermodified nucleoside mnm5s2U in tRNA |
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| Autoren: | Martine Roovers, Joël Caillet, Janusz Marek Bujnicki, Elzbieta Purta, Yamina Oudjama, Louis Droogmans, Katarzyna Kamińska |
| Weitere Verfasser: | Gauthier, Laurence |
| Quelle: | Proteins: Structure, Function, and Bioinformatics. 71:2076-2085 |
| Verlagsinformationen: | Wiley, 2008. |
| Publikationsjahr: | 2008 |
| Schlagwörter: | Models, Molecular, 0301 basic medicine, Secondary, Protein Folding, Sequence Homology, Ligands, Multienzyme Complexes -- analysis, Protein Structure, Secondary, RNA, Transfer, Computational Biology -- methods, Models, Escherichia coli Proteins -- chemistry, Conserved Sequence, Sequence Deletion, 0303 health sciences, Escherichia coli Proteins, Multienzyme Complexes -- genetics, Sciences bio-médicales et agricoles, Bacterial -- biosynthesis, Amino Acid, RNA, Bacterial, Flavin-Adenine Dinucleotide -- metabolism, Thiouridine -- metabolism, Escherichia coli Proteins -- genetics, Flavin-Adenine Dinucleotide, Sequence Analysis, Protein Binding, Protein Structure, Thiouridine -- analogs & derivatives, Static Electricity, Molecular Sequence Data, Methylation, Catalysis, Structure-Activity Relationship, 03 medical and health sciences, Transfer -- biosynthesis, Multienzyme Complexes, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Escherichia coli Proteins -- metabolism, Binding Sites, Genetic Complementation Test, Molecular, Computational Biology, Multienzyme Complexes -- chemistry, Protein Structure, Tertiary, Escherichia coli Proteins -- analysis, Amino Acid Substitution, RNA, Multienzyme Complexes -- metabolism, Tertiary |
| Beschreibung: | MnmC catalyses the last two steps in the biosynthesis of 5‐methylaminomethyl‐2‐thiouridine (mnm5s2U) in tRNA. Previously, we reported that this bifunctional enzyme is encoded by theyfcKopen reading frame in theEscherichia coliK12 genome. However, the mechanism of its activity, in particular the potential structural and functional dependence of the domains responsible for catalyzing the two modification reactions, remains unknown. With the aid of the protein fold‐recognition method, we constructed a structural model of MnmC in complex with the ligands and target nucleosides and studied the role of individual amino acids and entire domains by site‐directed and deletion mutagenesis, respectively. We found out that the N‐terminal domain contains residues responsible for binding of the S‐adenosylmethionine cofactor and catalyzing the methylation of nm5s2U to form mnm5s2U, while the C‐terminal domain contains residues responsible for binding of the FAD cofactor. Further, point mutants with compromised activity of either domain can complement each other to restore a fully functional enzyme. Thus, in the conserved fusion protein MnmC, the individual domains retain independence as enzymes. Interestingly, the N‐terminal domain is capable of independent folding, while the isolated C‐terminal domain is incapable of folding on its own, a situation similar to the one reported recently for the rRNA modification enzyme RsmC. Proteins, 2008. © 2008 Wiley‐Liss, Inc. |
| Publikationsart: | Article |
| Dateibeschreibung: | 1 full-text file(s): application/pdf |
| Sprache: | English |
| ISSN: | 1097-0134 0887-3585 |
| DOI: | 10.1002/prot.21918 |
| Zugangs-URL: | https://pubmed.ncbi.nlm.nih.gov/18186482 https://hal.science/hal-00310550v1 https://www.onlinelibrary.wiley.com/doi/abs/10.1002/prot.21918 https://www.ncbi.nlm.nih.gov/pubmed/18186482 https://www.onlinelibrary.wiley.com/doi/10.1002/prot.21918 https://difusion.ulb.ac.be/vufind/Record/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/50477/Details http://europepmc.org/abstract/MED/18186482 |
| Rights: | Wiley Online Library User Agreement |
| Dokumentencode: | edsair.doi.dedup.....1b08baa25a058fcf9477f0e24b01d5b5 |
| Datenbank: | OpenAIRE |
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Nájsť tento článok vo Web of Science | Abstract: | MnmC catalyses the last two steps in the biosynthesis of 5‐methylaminomethyl‐2‐thiouridine (mnm5s2U) in tRNA. Previously, we reported that this bifunctional enzyme is encoded by theyfcKopen reading frame in theEscherichia coliK12 genome. However, the mechanism of its activity, in particular the potential structural and functional dependence of the domains responsible for catalyzing the two modification reactions, remains unknown. With the aid of the protein fold‐recognition method, we constructed a structural model of MnmC in complex with the ligands and target nucleosides and studied the role of individual amino acids and entire domains by site‐directed and deletion mutagenesis, respectively. We found out that the N‐terminal domain contains residues responsible for binding of the S‐adenosylmethionine cofactor and catalyzing the methylation of nm5s2U to form mnm5s2U, while the C‐terminal domain contains residues responsible for binding of the FAD cofactor. Further, point mutants with compromised activity of either domain can complement each other to restore a fully functional enzyme. Thus, in the conserved fusion protein MnmC, the individual domains retain independence as enzymes. Interestingly, the N‐terminal domain is capable of independent folding, while the isolated C‐terminal domain is incapable of folding on its own, a situation similar to the one reported recently for the rRNA modification enzyme RsmC. Proteins, 2008. © 2008 Wiley‐Liss, Inc. |
|---|---|
| ISSN: | 10970134 08873585 |
| DOI: | 10.1002/prot.21918 |