Skip Header

You are using a version of browser that may not display all the features of this website. Please consider upgrading your browser.
Protein

Copper-exporting P-type ATPase

Gene

copA

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Copper-exporting P-type ATPase: Exports Cu+ from the cytoplasm to the periplasm (PubMed:10639134, PubMed:11167016, PubMed:11500054, PubMed:12351646). Binds 2 Cu+ ions per monomer, which are transferred to periplasmic copper chaperone CusF upon ATP hydrolysis (PubMed:24917681). In vitro an excess of CusF over CopA is required for efficient transfer (PubMed:24917681). May also be involved in silver export (PubMed:12351646, PubMed:12832075).6 Publications
Soluble copper chaperone CopA(Z): mRNA is subject to programmed ribosomal frameshifting which produces a cytoplasmic copper chaperone CopA(Z) that corresponds to the first HMA domain (PubMed:28107647). The soluble form is essential for cell survivial in the presence of CuSO4; in growth competition experiments between wild-type and a version that prevents expression of CopA(Z) after 50 generations the non-CopA(Z) version is nearly extinct (PubMed:28107647). The first HMA domain (residues 1-70) can be replaced by B.subtilis Cu chaperone CopZ (PubMed:25899340).2 Publications

Catalytic activityi

ATP + H2O + Cu+(Side 1) = ADP + phosphate + Cu+(Side 2).2 Publications

Enzyme regulationi

Copper-exporting P-type ATPase: Export is inhibited by vanadate (PubMed:10639134). Phosphorylation is inhibited by vanadate and sensitive to KOH and hydroxylamine; it is not inhibited by azide (PubMed:12351646). Phosphorylation is Cu+ not Cu2+-dependent (PubMed:12351646). ATPase activity is inhibited by bathocuproindisulfonate (BCDS), which chelates Cu+ but not Cu2+, and stimulated 3-4-fold by Cu+ (PubMed:12351646, PubMed:25899340). ATPase activity is inhibited by Cu2+ plus DTT or Ag+ (PubMed:12351646).3 Publications

Kineticsi

Export tested with Isoform Copper-exporting P-type ATPase.Curated
  1. KM=1.5 µM for copper1 Publication
  2. KM=0.5 mM for ATP1 Publication
  3. KM=1.48 µM for Cu+1 Publication
  4. KM=5.4 µM for Cu+1 Publication
  1. Vmax=0.19 µmol/min/mg enzyme (in the presence of CuCl2 and 1 mM DTT)1 Publication
  2. Vmax=1.64 µmol/h/mg enzyme for Cu+1 Publication

Sites

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Metal bindingi14CopperPROSITE-ProRule annotation1
Metal bindingi17CopperPROSITE-ProRule annotation1
Metal bindingi110CopperPROSITE-ProRule annotation1
Metal bindingi113CopperPROSITE-ProRule annotation1
Active sitei5234-aspartylphosphate intermediate1 Publication1
Metal bindingi720MagnesiumPROSITE-ProRule annotation1
Metal bindingi724MagnesiumPROSITE-ProRule annotation1

GO - Molecular functioni

  • ATPase activity Source: EcoliWiki
  • ATPase activity, coupled to transmembrane movement of ions, phosphorylative mechanism Source: EcoCyc
  • ATP binding Source: UniProtKB-KW
  • cation-transporting ATPase activity Source: InterPro
  • metal ion binding Source: UniProtKB-KW
  • silver ion transmembrane transporter activity Source: EcoCyc

GO - Biological processi

  • cellular response to copper ion Source: EcoCyc
  • cellular response to silver ion Source: EcoCyc
  • copper ion export Source: EcoCyc
  • copper ion transport Source: EcoliWiki
  • detoxification of copper ion Source: EcoCyc
  • silver ion transmembrane transport Source: EcoCyc

Keywordsi

Molecular functionChaperone, Hydrolase
Biological processCopper transport, Ion transport, Transport
LigandATP-binding, Copper, Magnesium, Metal-binding, Nucleotide-binding

Enzyme and pathway databases

BioCyciEcoCyc:G6260-MONOMER.
MetaCyc:G6260-MONOMER.
SABIO-RKiQ59385.

Protein family/group databases

TCDBi3.A.3.5.5. the p-type atpase (p-atpase) superfamily.

Names & Taxonomyi

Protein namesi
Recommended name:
Copper-exporting P-type ATPase1 Publication (EC:3.6.3.542 Publications)
Alternative name(s):
Copper-exporting P-type ATPase A
Cu(+)-exporting ATPase
Soluble copper chaperone CopA(Z)1 Publication
Gene namesi
Name:copA1 Publication
Synonyms:atcU1 Publication, f834, ybaR
Ordered Locus Names:b0484, JW0473
OrganismiEscherichia coli (strain K12)
Taxonomic identifieri83333 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacteralesEnterobacteriaceaeEscherichia
Proteomesi
  • UP000000318 Componenti: Chromosome
  • UP000000625 Componenti: Chromosome

Organism-specific databases

EcoGeneiEG13246. copA.

Subcellular locationi

Copper-exporting P-type ATPase :
  • Cell inner membrane 1 Publication1 Publication; Multi-pass membrane protein 1 Publication
Isoform Soluble copper chaperone CopA(Z) :

Topology

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Topological domaini2 – 186CytoplasmicCuratedAdd BLAST185
Transmembranei187 – 207HelicalSequence analysisAdd BLAST21
Topological domaini208 – 217Periplasmic; loop 11 Publication10
Transmembranei218 – 238HelicalSequence analysisAdd BLAST21
Topological domaini239 – 253CytoplasmicCuratedAdd BLAST15
Transmembranei254 – 274HelicalSequence analysisAdd BLAST21
Topological domaini275 – 283Periplasmic; loop 21 Publication9
Transmembranei284 – 304HelicalSequence analysisAdd BLAST21
Topological domaini305 – 437CytoplasmicCuratedAdd BLAST133
Transmembranei438 – 458HelicalSequence analysisAdd BLAST21
Topological domaini459 – 463Periplasmic; loop 3Curated5
Transmembranei464 – 484HelicalSequence analysisAdd BLAST21
Topological domaini485 – 778CytoplasmicCuratedAdd BLAST294
Transmembranei779 – 799HelicalSequence analysisAdd BLAST21
Topological domaini800Periplasmic; loop 41 Publication1
Transmembranei801 – 821HelicalSequence analysisAdd BLAST21
Topological domaini822 – 834Cytoplasmic1 PublicationAdd BLAST13

GO - Cellular componenti

  • cytoplasm Source: UniProtKB-SubCell
  • integral component of plasma membrane Source: EcoCyc
  • intrinsic component of membrane Source: EcoliWiki
  • membrane Source: UniProtKB
  • plasma membrane Source: EcoCyc

Keywords - Cellular componenti

Cell inner membrane, Cell membrane, Cytoplasm, Membrane

Pathology & Biotechi

Disruption phenotypei

Decreased resistance to Cu+ (PubMed:10639134, PubMed:11167016). No change in resistance to Zn2+ or Cd2+ (PubMed:10639134). Decreased resistance to AgNO3 (PubMed:12832075). Increased intracellular levels of Cu2+ (PubMed:11167016).3 Publications

Mutagenesis

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Mutagenesisi1 – 70Missing : Slight increase in CuSO(4)-stimulation of ATPase, no growth in CuSO(4). Grows when this protein fragment is provided in trans or if B.subtilis CopZ is present. 1 PublicationAdd BLAST70
Mutagenesisi3 – 113Missing : No resistance to CuSO(4), does not form phosphate intermediate. 1 PublicationAdd BLAST111
Mutagenesisi7 – 54Missing : Partial resistance to CuSO(4), forms phosphate intermediate. 1 PublicationAdd BLAST48
Mutagenesisi8 – 150Missing : Loss of growth in the presence of CuSO(4), loss of Cu efflux. 1 PublicationAdd BLAST143
Mutagenesisi14 – 17CGHC → AGHA: Wild-type growth in the presence of CuSO(4), no change in Cu efflux. Still forms phosphate intermediate; when associated with 110-A--A-113. 2 Publications4
Mutagenesisi14 – 17CGHC → SGHS: No change in CuSO(4)-stimulation of ATPase. When expressed as an isolated protein fragment (residues 1-70) does not restore growth to the 71-K--G-834 fragment. 1 Publication4
Mutagenesisi32 – 67EQADV…ASVSH → SRRMCLSLKRTLPGLPVQNS RSKPSNKRVMTHLYAN: Reduces -1 frameshifting efficiency about 2-fold in a 104 residue truncated construct. 1 PublicationAdd BLAST36
Mutagenesisi110 – 113CASC → AASA: Wild-type growth in the presence of CuSO(4), no change in Cu efflux. Still forms phosphate intermediate; when associated with 14-A--A-17. 2 Publications4
Mutagenesisi110 – 113CASC → SASS: Loss of CuSO(4)-stimulation of ATPase. When present in the 51-K--G-834 fragment growth in CuSO(4) is not restored by protein fragment 1-M--A-70. 1 Publication4
Mutagenesisi204M → A: Decreased transfer of Cu(+) to CusF, binds 2 Cu(+). 1 Publication1
Mutagenesisi207 – 210DNMM → AAAA: First half of periplasmic loop 1, transfers about 10% Cu(+) to CusF. 1 Publication4
Mutagenesisi212 – 216TADNR → AAANA: Second half of periplasmic loop 1, wild-type transfer of Cu(+) to CusF. 1 Publication5
Mutagenesisi273 – 277WPQWF → APQAA: First half of periplasmic loop 2, nearly wild-type transfer of Cu(+) to CusF. 1 Publication5
Mutagenesisi279 – 283MEARH → AAARA: Second half of periplasmic loop 2, wild-type transfer of Cu(+) to CusF. 1 Publication5
Mutagenesisi287E → A: Decreased transfer of Cu(+) to CusF. 1 Publication1
Mutagenesisi479C → A: Loss of copper resistance, transport and phosphoenzyme formation. 1 Publication1
Mutagenesisi481C → A or H: Loss of copper resistance, transport and phosphoenzyme formation. 1 Publication1
Mutagenesisi797 – 802WPFTGT → APFAGA: Periplasmic loop 4, nearly wild-type transfer of Cu(+) to CusF. 1 Publication6

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Initiator methionineiRemoved1 Publication
ChainiPRO_00000463202 – 834Copper-exporting P-type ATPaseAdd BLAST833

Keywords - PTMi

Phosphoprotein

Proteomic databases

PaxDbiQ59385.
PRIDEiQ59385.

Expressioni

Inductioni

Copper-exporting P-type ATPase: Induced by Cu2+ and Ag+ (at protein level) (PubMed:10639134). Transcriptionally regulated by CueR in response to Cu+ or Ag+ ions (PubMed:10639134, PubMed:11167016). Basal expression is low but unperturbed by disruption of cueR (PubMed:11167016).2 Publications

Interactioni

Subunit structurei

Copper-exporting P-type ATPase interacts with apo-periplasmic copper chaperone CusF; when CusF is precharged with copper it binds very little CopA. The periplasmic loops of CopA, especially the first half of loop 1, play a large role in binding to CusF (PubMed:24917681).1 Publication

Protein-protein interaction databases

BioGridi4261336. 9 interactors.
IntActiQ59385. 6 interactors.
STRINGi316385.ECDH10B_0441.

Structurei

3D structure databases

ProteinModelPortaliQ59385.
SMRiQ59385.
ModBaseiSearch...
MobiDBiSearch...

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Domaini4 – 65HMA 1PROSITE-ProRule annotationAdd BLAST62
Domaini100 – 163HMA 2PROSITE-ProRule annotationAdd BLAST64

Motif

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Motifi14 – 17CXXC motif 11 Publication4
Motifi110 – 113CXXC motif 21 Publication4

Domaini

The N-terminal domain (exact residues are not given in the paper) is not required for Cu+-binding (when deleted KM for Cu+ binding is 1.32 µM) nor for ATPase activity, binds 2 Cu+/monomer (PubMed:24917681). Contradictory results give a considerable decrease in Cu affinity when residues 1-150 are deleted (KM=31.9 µM for Cu+) (PubMed:25899340). The first of 2 N-terminal heavy metal-binding domains (HMA 1, approximately residues 1-70, equivalent to CopA(Z)) has a 5-fold higher affinity for Cu+ than HMA 2 (residues 71-150) and as a protein fragment can transfer Cu+ to the ATPase fragment (residues 151-834), suggesting it has a Cu-chaperone function (PubMed:25899340). HMA 2 tranfers Cu+ to HMA 1 but the opposite reaction does not occur in vitro (PubMed:25899340). The HMA 1 fragment complements growth defects in trans, but if its CXXC motif is mutated, or if the remaining CXXC motif in HMA2 is mutated, complementation no longer occurs, showing the 2 HMA domains have different functions (PubMed:25899340). The periplasmic loops of CopA, especially the first half of loop 1, play a large role in binding to CusF (PubMed:24917681). Contradictory results between the various in vitro studies may be due to different levels of protein expression or reconstitution (Probable).Curated2 Publications

Sequence similaritiesi

Keywords - Domaini

Repeat, Transmembrane, Transmembrane helix

Phylogenomic databases

eggNOGiENOG4105C59. Bacteria.
COG2217. LUCA.
HOGENOMiHOG000250397.
InParanoidiQ59385.
KOiK17686.
PhylomeDBiQ59385.

Family and domain databases

CDDicd00371. HMA. 2 hits.
Gene3Di3.40.1110.10. 1 hit.
3.40.50.1000. 1 hit.
InterProiView protein in InterPro
IPR023299. ATPase_P-typ_cyto_dom_N.
IPR018303. ATPase_P-typ_P_site.
IPR023298. ATPase_P-typ_TM_dom_sf.
IPR008250. ATPase_P-typ_transduc_dom_A_sf.
IPR036412. HAD-like_sf.
IPR023214. HAD_sf.
IPR017969. Heavy-metal-associated_CS.
IPR006121. HMA_dom.
IPR036163. HMA_dom_sf.
IPR027256. P-typ_ATPase_IB.
IPR001757. P_typ_ATPase.
PfamiView protein in Pfam
PF00403. HMA. 2 hits.
PRINTSiPR00120. HATPASE.
SUPFAMiSSF55008. SSF55008. 2 hits.
SSF56784. SSF56784. 2 hits.
SSF81653. SSF81653. 1 hit.
SSF81665. SSF81665. 3 hits.
TIGRFAMsiTIGR01525. ATPase-IB_hvy. 1 hit.
TIGR01494. ATPase_P-type. 1 hit.
PROSITEiView protein in PROSITE
PS00154. ATPASE_E1_E2. 1 hit.
PS01047. HMA_1. 1 hit.
PS50846. HMA_2. 2 hits.

Sequences (2)i

Sequence statusi: Complete.

Sequence processingi: The displayed sequence is further processed into a mature form.

This entry describes 2 isoformsi produced by ribosomal frameshifting. AlignAdd to basket

Isoform Copper-exporting P-type ATPase (identifier: Q59385-1) [UniParc]FASTAAdd to basket

This isoform has been chosen as the 'canonical' sequence. All positional information in this entry refers to it. This is also the sequence that appears in the downloadable versions of the entry.

« Hide

        10         20         30         40         50
MSQTIDLTLD GLSCGHCVKR VKESLEQRPD VEQADVSITE AHVTGTASAE
60 70 80 90 100
QLIETIKQAG YDASVSHPKA KPLAESSIPS EALTAVSEAL PAATADDDDS
110 120 130 140 150
QQLLLSGMSC ASCVTRVQNA LQSVPGVTQA RVNLAERTAL VMGSASPQDL
160 170 180 190 200
VQAVEKAGYG AEAIEDDAKR RERQQETAVA TMKRFRWQAI VALAVGIPVM
210 220 230 240 250
VWGMIGDNMM VTADNRSLWL VIGLITLAVM VFAGGHFYRS AWKSLLNGAA
260 270 280 290 300
TMDTLVALGT GVAWLYSMSV NLWPQWFPME ARHLYYEASA MIIGLINLGH
310 320 330 340 350
MLEARARQRS SKALEKLLDL TPPTARLVTD EGEKSVPLAE VQPGMLLRLT
360 370 380 390 400
TGDRVPVDGE ITQGEAWLDE AMLTGEPIPQ QKGEGDSVHA GTVVQDGSVL
410 420 430 440 450
FRASAVGSHT TLSRIIRMVR QAQSSKPEIG QLADKISAVF VPVVVVIALV
460 470 480 490 500
SAAIWYFFGP APQIVYTLVI ATTVLIIACP CALGLATPMS IISGVGRAAE
510 520 530 540 550
FGVLVRDADA LQRASTLDTV VFDKTGTLTE GKPQVVAVKT FADVDEAQAL
560 570 580 590 600
RLAAALEQGS SHPLARAILD KAGDMQLPQV NGFRTLRGLG VSGEAEGHAL
610 620 630 640 650
LLGNQALLNE QQVGTKAIEA EITAQASQGA TPVLLAVDGK AVALLAVRDP
660 670 680 690 700
LRSDSVAALQ RLHKAGYRLV MLTGDNPTTA NAIAKEAGID EVIAGVLPDG
710 720 730 740 750
KAEAIKHLQS EGRQVAMVGD GINDAPALAQ ADVGIAMGGG SDVAIETAAI
760 770 780 790 800
TLMRHSLMGV ADALAISRAT LHNMKQNLLG AFIYNSIGIP VAAGILWPFT
810 820 830
GTLLNPVVAG AAMALSSITV VSNANRLLRF KPKE
Length:834
Mass (Da):87,873
Last modified:January 23, 2007 - v4
Checksum:iCF84A18FE208E6F6
GO
Isoform Soluble copper chaperone CopA(Z) (identifier: Q59385-2) [UniParc]FASTAAdd to basket

The sequence of this isoform differs from the canonical sequence as follows:
     70-834: AKPLAESSIP...NRLLRFKPKE → G

Note: Expression of the CopA(Z) soluble copper chaperone isoform requires a -1 programmed ribosomal frameshift (PRF) at the 70th codon, promoted by a nucleotide 'slippery sequence'. Silent mutations in the 'slippery sequence' abrogate expression of CopA(Z) but still allow expression of the full length protein. Both the mRNA secondary structure (a possible pseudoknot just downstream of the slippage site) and the sequence of the protein in the ribosomal exit tunnel modulate the efficiency of the -1 PRF (PubMed:28107647).1 Publication
Show »
Length:70
Mass (Da):7,481
Checksum:iF0370BB67BCFDC7A
GO

Experimental Info

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Sequence conflicti162 – 181EAIED…TAVAT → KRLKMTLNAASASKKPPSLA in AAB02268 (Ref. 1) CuratedAdd BLAST20
Sequence conflicti508A → R in AAB02268 (Ref. 1) Curated1
Sequence conflicti576Q → R in AAB02268 (Ref. 1) Curated1

Alternative sequence

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Alternative sequenceiVSP_05917670 – 834AKPLA…FKPKE → G in isoform Soluble copper chaperone CopA(Z). 1 PublicationAdd BLAST765

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
U58330 Genomic DNA. Translation: AAB02268.1.
U82664 Genomic DNA. Translation: AAB40238.1.
U00096 Genomic DNA. Translation: AAC73586.1.
AP009048 Genomic DNA. Translation: BAE76263.1.
PIRiC64779.
RefSeqiNP_415017.1. NC_000913.3.
WP_000083955.1. NZ_LN832404.1.

Genome annotation databases

EnsemblBacteriaiAAC73586; AAC73586; b0484.
BAE76263; BAE76263; BAE76263.
GeneIDi946106.
KEGGiecj:JW0473.
eco:b0484.
PATRICifig|1411691.4.peg.1792.

Keywords - Coding sequence diversityi

Ribosomal frameshifting

Similar proteinsi

Entry informationi

Entry nameiCOPA_ECOLI
AccessioniPrimary (citable) accession number: Q59385
Secondary accession number(s): P78245, Q2MBU3
Entry historyiIntegrated into UniProtKB/Swiss-Prot: November 1, 1997
Last sequence update: January 23, 2007
Last modified: November 22, 2017
This is version 160 of the entry and version 4 of the sequence. See complete history.
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

Complete proteome, Direct protein sequencing, Reference proteome

Documents

  1. Escherichia coli
    Escherichia coli (strain K12): entries and cross-references to EcoGene
  2. SIMILARITY comments
    Index of protein domains and families