| IED ID | IndEnz0015000101 |
| Enzyme Type ID | laccase000101 |
| Protein Name |
Multicopper oxidase CueO MCO EC 1.16.3.- Blue copper oxidase CueO Copper efflux oxidase Cu efflux oxidase Cuprous oxidase |
| Gene Name | cueO yacK b0123 JW0119 |
| Organism | Escherichia coli (strain K12) |
| Taxonomic Lineage | cellular organisms Bacteria Proteobacteria Gammaproteobacteria Enterobacterales Enterobacteriaceae Escherichia Escherichia coli Escherichia coli (strain K12) |
| Enzyme Sequence | MQRRDFLKYSVALGVASALPLWSRAVFAAERPTLPIPDLLTTDARNRIQLTIGAGQSTFGGKTATTWGYNGNLLGPAVKLQRGKAVTVDIYNQLTEETTLHWHGLEVPGEVDGGPQGIIPPGGKRSVTLNVDQPAATCWFHPHQHGKTGRQVAMGLAGLVVIEDDEILKLMLPKQWGIDDVPVIVQDKKFSADGQIDYQLDVMTAAVGWFGDTLLTNGAIYPQHAAPRGWLRLRLLNGCNARSLNFATSDNRPLYVIASDGGLLPEPVKVSELPVLMGERFEVLVEVNDNKPFDLVTLPVSQMGMAIAPFDKPHPVMRIQPIAISASGALPDTLSSLPALPSLEGLTVRKLQLSMDPMLDMMGMQMLMEKYGDQAMAGMDHSQMMGHMGHGNMNHMNHGGKFDFHHANKINGQAFDMNKPMFAAAKGQYERWVISGVGDMMLHPFHIHGTQFRILSENGKPPAAHRAGWKDTVKVEGNVSEVLVKFNHDAPKEHAYMAHCHLLEHEDTGMMLGFTV |
| Enzyme Length | 516 |
| Uniprot Accession Number | P36649 |
| Absorption | |
| Active Site | |
| Activity Regulation | ACTIVITY REGULATION: Ferroxidase and phenoloxidase activities are enhanced considerably in the presence of excess copper ions (PubMed:11466290, PubMed:11527384, PubMed:11867755, PubMed:15516598, PubMed:17804014). A labile regulatory copper ion near the T1 copper site is important for the copper associated activation of enzyme activity (PubMed:12794077). Ag(+) acts as a potent inhibitor of oxidase activity by binding at Cu(+) binding sites, blocking Cu(+) substrate binding and oxidation (PubMed:21903583). pPD oxidase activity is strongly inhibited by sodium azide, an inhibitor of the electron transfer (PubMed:11527384). {ECO:0000269|PubMed:11466290, ECO:0000269|PubMed:11527384, ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:15516598, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=4 Cu(+) + 4 H(+) + O2 = 4 Cu(2+) + 2 H2O; Xref=Rhea:RHEA:30083, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:29036, ChEBI:CHEBI:49552; Evidence={ECO:0000269|PubMed:15516598, ECO:0000269|PubMed:20088522};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:30084; Evidence={ECO:0000269|PubMed:15516598, ECO:0000269|PubMed:20088522}; |
| DNA Binding | |
| EC Number | 1.16.3.- |
| Enzyme Function | FUNCTION: Multicopper oxidase involved in copper homeostasis and copper tolerance under aerobic conditions (PubMed:11222619, PubMed:11399769, PubMed:11527384, PubMed:15516598). Is responsible for the oxidation of Cu(+) to the less harmful Cu(2+) in the periplasm, thereby preventing Cu(+) from entering the cytoplasm (PubMed:15516598, PubMed:20088522, PubMed:25679350). Probably primarily functions as a cuprous oxidase in vivo (PubMed:20088522). {ECO:0000269|PubMed:11222619, ECO:0000269|PubMed:11399769, ECO:0000269|PubMed:11527384, ECO:0000269|PubMed:15516598, ECO:0000269|PubMed:20088522, ECO:0000269|PubMed:25679350}.; FUNCTION: In vitro, in the presence of excess copper ions, exhibits ferroxidase and phenoloxidase activities (PubMed:11466290, PubMed:11527384, PubMed:11867755, PubMed:15317788, PubMed:15516598, PubMed:17804014, PubMed:25679350). Fe(2+) is an excellent substrate in the presence of excess Cu(2+), but is inactive in the absence of Cu(2+) (PubMed:15516598, PubMed:17804014). Oxidizes the phenolate iron siderophores enterobactin, 2,3-dihydroxybenzoate (2,3-DHB) and 3-hydroxyanthranilate (3-HAA) (PubMed:11466290, PubMed:15317788). Oxidation and thus inactivation of enterobactin could protect cells from the interaction of enterobactin with copper and play a central role as an interface between copper detoxification and iron homeostasis (PubMed:11466290, PubMed:15317788). Also oxidizes a variety of phenolic model substrates, including 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) (ABTS), p-phenylenediamine (pPD), 2,6-dimethoxyphenol (2,6-DMP) and 3,4-dihydroxybenzoic acid (3,4-DHB) (PubMed:11466290, PubMed:11527384, PubMed:17804014, PubMed:25679350). {ECO:0000269|PubMed:11466290, ECO:0000269|PubMed:11527384, ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:15317788, ECO:0000269|PubMed:15516598, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:25679350}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 55 degrees Celsius with 2,6-DMP as substrate. {ECO:0000269|PubMed:11867755}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 6.5 with 2,6-DMP as substrate (PubMed:11867755). The pH dependence of the reaction as monitored by oxygen consumption shows a broad activity peak between pH 5 and 6 and a second activity peak at pH 8 (PubMed:12794077). {ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077}; |
| Pathway | |
| nucleotide Binding | |
| Features | Beta strand (31); Chain (1); Domain (3); Helix (14); Metal binding (11); Mutagenesis (12); Region (1); Signal peptide (1); Turn (5) |
| Keywords | 3D-structure;Copper;Direct protein sequencing;Metal-binding;Oxidoreductase;Periplasm;Reference proteome;Repeat;Signal |
| Interact With | |
| Induction | INDUCTION: By CueR, in response to increasing copper concentrations. {ECO:0000269|PubMed:10915804}. |
| Subcellular Location | SUBCELLULAR LOCATION: Periplasm {ECO:0000269|PubMed:10766774, ECO:0000269|PubMed:11399769, ECO:0000269|PubMed:11527384, ECO:0000269|PubMed:27129241}. Note=Exported via the Tat pathway (PubMed:10766774, PubMed:17218314, PubMed:27129241). Cytoplasmic CueO does not contain copper, even under copper stress conditions, and is transported as an apo-protein to the periplasm. Periplasmic CueO is readily activated by the addition of copper ions in vitro or under copper stress conditions in vivo (PubMed:27129241). Can also be exported by the Sec system (PubMed:17218314). {ECO:0000269|PubMed:10766774, ECO:0000269|PubMed:17218314, ECO:0000269|PubMed:27129241}. |
| Modified Residue | |
| Post Translational Modification | PTM: Exported by the Tat system (PubMed:10766774, PubMed:17218314, PubMed:27129241). The position of the signal peptide cleavage has been experimentally proven (PubMed:9298646). {ECO:0000269|PubMed:10766774, ECO:0000269|PubMed:17218314, ECO:0000269|PubMed:27129241, ECO:0000269|PubMed:9298646}. |
| Signal Peptide | SIGNAL 1..28; /note="Tat-type signal"; /evidence="ECO:0000255|PROSITE-ProRule:PRU00648, ECO:0000269|PubMed:9298646" |
| Structure 3D | X-ray crystallography (39) |
| Cross Reference PDB | 1KV7; 1N68; 1PF3; 2FQD; 2FQE; 2FQF; 2FQG; 2YXV; 2YXW; 3NSC; 3NSD; 3NSF; 3NSY; 3NT0; 3OD3; 3PAU; 3PAV; 3QQX; 3UAA; 3UAB; 3UAC; 3UAD; 3UAE; 4E9Q; 4E9R; 4E9S; 4E9T; 4EF3; 4HAK; 4HAL; 4NER; 5B7E; 5B7F; 5B7M; 5YS1; 5YS5; 6IM7; 6IM8; 6IM9; |
| Mapped Pubmed ID | 12142427; 16606699; 22250058; 27710945; |
| Motif | |
| Gene Encoded By | |
| Mass | 56,556 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=165 uM for Cu(+) (at pH 5.0, in the absence of added Cu(2+)) {ECO:0000269|PubMed:15516598}; KM=169 uM for Cu(+) (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:15516598}; KM=90 uM for Cu(+) (at pH 7.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:15516598}; KM=129 uM for Fe(2+) (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:15516598}; KM=70 uM for Fe(2+) (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=40 uM for enterobactin (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=1.5 uM for ferric enterobactin (at pH 6.5, in the presence of 0.5 mM Cu(2+)) {ECO:0000269|PubMed:15317788}; KM=290 uM for 2,3-DHB (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=690 uM for 3-HAA (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=2500 uM for ABTS (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=3200 uM for pPD (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=2120 uM for 2,6-DMP (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; KM=70.5 uM for 2,6-DMP (at pH 6.5, in the presence of 250 uM Cu(2+)) {ECO:0000269|PubMed:11867755}; KM=430 uM for 3,4-DHB (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=41.0 umol/min/mg enzyme with Fe(2+) as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=9.70 umol/min/mg enzyme with enterobactin as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=3.80 umol/min/mg enzyme with 2,3-DHB as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=63.90 umol/min/mg enzyme with 3-HAA as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=72.10 umol/min/mg enzyme with ABTS as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=95.90 umol/min/mg enzyme with pPD as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=91.70 umol/min/mg enzyme with 2,6-DMP as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Vmax=7.20 umol/min/mg enzyme with 3,4-DHB as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) {ECO:0000269|PubMed:11466290}; Note=kcat is 914 min(-1) with Cu(+) as substrate (at pH 5.0, in the absence of added Cu(2+)) (PubMed:15516598). kcat is 651 min(-1) with Cu(+) as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) (PubMed:15516598). kcat is 57 min(-1) with Cu(+) as substrate (at pH 7.0, in the presence of 1 mM Cu(2+)) (PubMed:15516598). kcat is 215 min(-1) with Fe(2+) as substrate (at pH 5.0, in the presence of 1 mM Cu(2+)) (PubMed:15516598). kcat is 120 sec(-1) with 2,6-DMP as substrate (PubMed:11867755). {ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:15516598}; |
| Metal Binding | METAL 101; /note="Copper 1; via tele nitrogen; type 2"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 103; /note="Copper 2; via pros nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 141; /note="Copper 2; via tele nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 143; /note="Copper 3; via tele nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 443; /note="Copper 4; via pros nitrogen; type 1"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 446; /note="Copper 1; via tele nitrogen; type 2"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 448; /note="Copper 3; via tele nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 499; /note="Copper 3; via tele nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 500; /note="Copper 4; type 1"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 501; /note="Copper 2; via tele nitrogen; type 3"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW"; METAL 505; /note="Copper 4; via pros nitrogen; type 1"; /evidence="ECO:0000269|PubMed:11867755, ECO:0000269|PubMed:12794077, ECO:0000269|PubMed:17217912, ECO:0000269|PubMed:17804014, ECO:0000269|PubMed:21903583, ECO:0000269|PubMed:24598746, ECO:0007744|PDB:1KV7, ECO:0007744|PDB:1N68, ECO:0007744|PDB:1PF3, ECO:0007744|PDB:2FQD, ECO:0007744|PDB:2FQE, ECO:0007744|PDB:2FQF, ECO:0007744|PDB:2FQG, ECO:0007744|PDB:2YXV, ECO:0007744|PDB:2YXW" |
| Rhea ID | RHEA:30083; RHEA:30084 |
| Cross Reference Brenda |