moeA (PP_2123) encodes the soluble molybdopterin molybdotransferase of Pseudomonas putida KT2440. By family membership with Escherichia coli MoeA it inserts molybdate into adenylylated molybdopterin (MPT-AMP) with release of AMP, forming Mo-molybdopterin, the metal-insertion step of molybdenum cofactor biosynthesis that precedes the optional nucleotide additions made by MobA. The protein has the full MoeA four-domain architecture (MoeA N-terminal/linker domain, MoaB/Mog-like domain and MoeA C-terminal domain) and a Mg2+ cofactor; it has not been characterised directly in KT2440.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005737 cytoplasm | IEA GO_REF:0000118 | MODIFY | Summary: TreeGrafter transfer of cytoplasm from PANTHER PTHR10192 (node PTN002229918) for this soluble molybdopterin molybdotransferase. Reason: True but less specific than the evidence supports. The same PANTHER transfer also gives cytosol (GO:0005829), a part of the cytoplasm, and the characterised E. coli ortholog MoeA is found in the cytosolic fraction; so the row is refined to cytosol, as for ECOLI/moeA. This revises the earlier KEEP_AS_NON_CORE from the TreeGrafter re-review, which argued that redundancy is not over-annotation; that is right, but a correct-but-general term is a MODIFY to the supported descendant, not a non-core keep. Proposed replacements: cytosol Supporting Evidence: PMID:18304323 we identified 1103 proteins from the cytosolic fraction of the Escherichia coli file:PSEPK/moeA/moeA-uniprot.txt PANTHER; PTHR10192; MOLYBDOPTERIN BIOSYNTHESIS PROTEIN; 1. |
| GO:0005829 cytosol | IEA GO_REF:0000118 | KEEP AS NON CORE | Summary: Cytosol is an appropriate specific location for MoeA. Reason: The molybdate-insertion reaction occurs in the soluble cofactor pathway; the protein has no membrane or export features, and E. coli MoeA is cytosolic. Location is peripheral to function, as in ECOLI/moeA. Supporting Evidence: PMID:18304323 we identified 1103 proteins from the cytosolic fraction of the Escherichia coli |
| GO:0006777 Mo-molybdopterin cofactor biosynthetic process | IEA GO_REF:0000120 | ACCEPT | Summary: MoeA directly forms Mo-molybdopterin during cofactor biosynthesis. Reason: The curated RuleBase function assigns the Mo-insertion reaction, and the E. coli ortholog performs it with purified components; MoeA does work in the process. Same action as ECOLI/moeA. Supporting Evidence: file:PSEPK/moeA/moeA-uniprot.txt FUNCTION: Catalyzes the insertion of molybdate into adenylated molybdopterin with the concomitant release of AMP. PMID:15632135 The results presented here demonstrate that MoeA is responsible for mediating molybdenum ligation to molybdopterin |
| GO:0032324 molybdopterin cofactor biosynthetic process | IEA GO_REF:0000120 | MODIFY | Summary: The general molybdopterin cofactor process is correct but less precise than the Mo-molybdopterin cofactor process the gene also carries. Reason: Correct but general, so MODIFY rather than MARK_AS_OVER_ANNOTATED (the earlier draft action). GO:0032324 covers Mo- and W-molybdopterin cofactors; MoeA in KT2440, a molybdenum-using bacterium, forms Mo-molybdopterin, and the gene already carries the descendant GO:0006777. Same action as ECOLI/moeA. Proposed replacements: Mo-molybdopterin cofactor biosynthetic process Supporting Evidence: file:PSEPK/moeA/moeA-uniprot.txt FUNCTION: Catalyzes the insertion of molybdate into adenylated molybdopterin with the concomitant release of AMP. |
| GO:0046872 metal ion binding | IEA GO_REF:0000104 | MODIFY | Summary: The metal bound by MoeA is a Mg2+ cofactor (one per subunit in the E. coli structure). Reason: Correct but general. The curated RuleBase cofactor line names Mg2+, and the E. coli MoeA crystal structure binds one Mg2+ per subunit, so the specific descendant GO:0000287 magnesium ion binding is supported. Molybdate is a substrate, not a bound metal ion cofactor. The earlier draft kept the generic term as non-core while arguing it was less informative; that action did not match the reason. Proposed replacements: magnesium ion binding Supporting Evidence: file:PSEPK/moeA/moeA-uniprot.txt Name=Mg(2+); Xref=ChEBI:CHEBI:18420; file:ECOLI/moeA/moeA-uniprot.txt Binds 1 Mg(2+) ion per subunit. |
| GO:0061599 molybdopterin molybdotransferase activity | IEA GO_REF:0000120 | ACCEPT | Summary: This is the exact core molecular function of MoeA. Reason: The curated UniRule (RuleBase RU365090) function statement assigns insertion of molybdate into adenylated molybdopterin with release of AMP, and the protein carries the full MoeA domain architecture (InterPro MoeA-like; Pfam MoeA_N, MoCF_biosynth and MoeA_C). The CATALYTIC ACTIVITY line itself rests only on ARBA and is not used. The activity is shown for the E. coli ortholog with purified components (PMID:15632135); there is no KT2440 enzyme assay. Supporting Evidence: file:PSEPK/moeA/moeA-deep-research-openscientist.md > **adenylyl-molybdopterin (MPT-AMP) + molybdate (MoO₄²⁻) → Mo-molybdopterin (Moco) + AMP + H⁺** (Mg²⁺-dependent) file:PSEPK/moeA/moeA-uniprot.txt Catalyzes the insertion of molybdate into adenylated file:PSEPK/moeA/moeA-uniprot.txt InterPro; IPR038987; MoeA-like. file:PSEPK/moeA/moeA-uniprot.txt Pfam; PF03453; MoeA_N; 1. PMID:15632135 The results presented here demonstrate that MoeA is responsible for mediating molybdenum ligation to molybdopterin |
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Download this section (compressed HTML)Q: Which KT2440 protein supplies the preceding MPT adenylyltransferase activity required by MoeA?
Experiment: Reconstitute Mo-MPT formation with Q88L14 and candidate MPT-adenylylating proteins, measuring adenylyl-MPT, AMP release, and Mo-MPT formation.
Type: biochemical pathway reconstitution
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