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P11439 (TOXA_PSEAE) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 119. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (1) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Exotoxin A

Short name=ETA
EC=2.4.2.36
Alternative name(s):
NAD(+)--diphthamide ADP-ribosyltransferase
PE
Gene names
Name:eta
Ordered Locus Names:PA1148
OrganismPseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) [Reference proteome] [HAMAP]
Taxonomic identifier208964 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaPseudomonadalesPseudomonadaceaePseudomonas

Protein attributes

Sequence length638 AA.
Sequence statusComplete.
Sequence processingThe displayed sequence is further processed into a mature form.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

An NAD-dependent ADP-ribosyltransferase (ADPRT). Catalyzes the transfer of the ADP ribosyl moiety of oxidized NAD (NAD+) onto eukaryotic elongation factor 2 (eEF-2) thus arresting protein synthesis. Has an LD50 of 65 ng/ml against the human lung epithelial cell line C38. Ref.8

Catalytic activity

NAD+ + diphthamide-[translation elongation factor 2] = nicotinamide + N-(ADP-D-ribosyl)diphthamide-[translation elongation factor 2].

Enzyme regulation

Inhibited by 1,8-naphthalimide (NAP) as well as a number of poly(ADP-ribose) polymerase inhibitors and other compounds. Ref.9

Post-translational modification

The 8 cysteines participate in intrachain disulfide bonds.

Biophysicochemical properties

Kinetic parameters:

For ADP-ribosyltransferase activity of the catalytic fragment 399-605.

KM=121 µM for NAD

Ontologies

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Signal peptide1 – 2525 Ref.1
Chain26 – 638613Exotoxin A
PRO_0000019365

Regions

Nucleotide binding465 – 4673NAD
Nucleotide binding479 – 4857NAD
Region26 – 277252IA (required for target cell recognition)
Region278 – 389112II (required for translocation in target cell cytoplasm)
Region390 – 42940IB
Region430 – 638209III (required for ADP-ribosyl activity)

Sites

Active site5781 Ref.3
Binding site4741NAD
Binding site5781NAD

Amino acid modifications

Disulfide bond290 ↔ 312 Ref.7

Experimental info

Mutagenesis821K → E: Loss of toxicity, no binding to LRP1 receptor. Ref.6
Mutagenesis4811R → H: 52-fold decrease in ADPRT activity, 55-fold reduction in GH activity, 31-fold increase in dissociation constant for NAD(+). Ref.12
Mutagenesis5711E → A: 833-fold decrease in ADPRT activity, 4-fold reduction in GH activity, 2-fold increase in dissociation constant for NAD(+). Ref.12
Mutagenesis5761R → A: 2-fold decrease in ADPRT activity, 14-fold reduction in GH activity, 3-fold increase in dissociation constant for NAD(+). Ref.12
Mutagenesis5781E → A: 666-fold decrease in ADPRT activity, 14-fold reduction in GH activity, 2-fold increase in dissociation constant for NAD(+), loss of toxicity against mouse cells. Ref.8 Ref.12
Sequence conflict41T → I in AAB59097. Ref.1
Sequence conflict221F → S in AAB59097. Ref.1
Sequence conflict2041A → T in AAB59097. Ref.1
Sequence conflict3891S → N in AAB59097. Ref.1
Sequence conflict4321I → V in AAB59097. Ref.1
Sequence conflict5401G → S in AAB59097. Ref.1

Secondary structure

........................................................................................................................ 638
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P11439 [UniParc].

Last modified January 11, 2001. Version 2.
Checksum: 7B9AAD56A27C700A

FASTA63869,284
        10         20         30         40         50         60 
MHLTPHWIPL VASLGLLAGG SFASAAEEAF DLWNECAKAC VLDLKDGVRS SRMSVDPAIA 

        70         80         90        100        110        120 
DTNGQGVLHY SMVLEGGNDA LKLAIDNALS ITSDGLTIRL EGGVEPNKPV RYSYTRQARG 

       130        140        150        160        170        180 
SWSLNWLVPI GHEKPSNIKV FIHELNAGNQ LSHMSPIYTI EMGDELLAKL ARDATFFVRA 

       190        200        210        220        230        240 
HESNEMQPTL AISHAGVSVV MAQAQPRREK RWSEWASGKV LCLLDPLDGV YNYLAQQRCN 

       250        260        270        280        290        300 
LDDTWEGKIY RVLAGNPAKH DLDIKPTVIS HRLHFPEGGS LAALTAHQAC HLPLETFTRH 

       310        320        330        340        350        360 
RQPRGWEQLE QCGYPVQRLV ALYLAARLSW NQVDQVIRNA LASPGSGGDL GEAIREQPEQ 

       370        380        390        400        410        420 
ARLALTLAAA ESERFVRQGT GNDEAGAASA DVVSLTCPVA AGECAGPADS GDALLERNYP 

       430        440        450        460        470        480 
TGAEFLGDGG DISFSTRGTQ NWTVERLLQA HRQLEERGYV FVGYHGTFLE AAQSIVFGGV 

       490        500        510        520        530        540 
RARSQDLDAI WRGFYIAGDP ALAYGYAQDQ EPDARGRIRN GALLRVYVPR SSLPGFYRTG 

       550        560        570        580        590        600 
LTLAAPEAAG EVERLIGHPL PLRLDAITGP EEEGGRLETI LGWPLAERTV VIPSAIPTDP 

       610        620        630 
RNVGGDLDPS SIPDKEQAIS ALPDYASQPG KPPREDLK 

« Hide

References

« Hide 'large scale' references
[1]"Cloning, nucleotide sequence, and expression in Escherichia coli of the exotoxin A structural gene of Pseudomonas aeruginosa."
Gray G.L., Smith D.H., Baldridge J.S., Harkins R.N., Vasil M.L., Chen E.Y., Heyneker H.L.
Proc. Natl. Acad. Sci. U.S.A. 81:2645-2649(1984) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 26-53.
[2]"Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen."
Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P., Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M., Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y., Brody L.L., Coulter S.N., Folger K.R. expand/collapse author list , Kas A., Larbig K., Lim R.M., Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J., Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.
Nature 406:959-964(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228.
[3]"Active site of Pseudomonas aeruginosa exotoxin A. Glutamic acid 553 is photolabeled by NAD and shows functional homology with glutamic acid 148 of diphtheria toxin."
Carroll S.F., Collier R.J.
J. Biol. Chem. 262:8707-8711(1987) [PubMed] [Europe PMC] [Abstract]
Cited for: ACTIVE SITE.
[4]"Mutagenesis of Pseudomonas exotoxin in identification of sequences responsible for the animal toxicity."
Chaudhary V.K., Jinno Y., Galo M.G., Fitzgerald D., Pastan I.
J. Biol. Chem. 265:16306-16310(1990) [PubMed] [Europe PMC] [Abstract]
Cited for: DOMAINS.
[5]"Biochemical and immunochemical studies of proteolytic fragments of exotoxin A from Pseudomonas aeruginosa."
Bourdenet S., Vacheron M.-J., Guinand M., Michel G., Arminjon F.
Eur. J. Biochem. 192:379-385(1990) [PubMed] [Europe PMC] [Abstract]
Cited for: DOMAINS.
[6]"The alpha 2-macroglobulin receptor/low density lipoprotein receptor-related protein binds and internalizes Pseudomonas exotoxin A."
Kounnas M.Z., Morris R.E., Thompson M.R., FitzGerald D.J., Strickland D.K., Saelinger C.B.
J. Biol. Chem. 267:12420-12423(1992) [PubMed] [Europe PMC] [Abstract]
Cited for: RECEPTOR-BINDING, MUTAGENESIS OF LYS-82.
[7]"Reduction of furin-nicked Pseudomonas exotoxin A: an unfolding story."
McKee M.L., FitzGerald D.J.
Biochemistry 38:16507-16513(1999) [PubMed] [Europe PMC] [Abstract]
Cited for: DISULFIDE BOND.
[8]"Cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae."
Jorgensen R., Purdy A.E., Fieldhouse R.J., Kimber M.S., Bartlett D.H., Merrill A.R.
J. Biol. Chem. 283:10671-10678(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION AS A TOXIN, FUNCTION AS A GLYCOHYDROLASE, FUNCTION AS AN ADP-RIBOSYLTRANSFERASE, MUTAGENESIS OF GLU-578.
[9]"Newly discovered and characterized antivirulence compounds inhibit bacterial mono-ADP-ribosyltransferase toxins."
Turgeon Z., Jorgensen R., Visschedyk D., Edwards P.R., Legree S., McGregor C., Fieldhouse R.J., Mangroo D., Schapira M., Merrill A.R.
Antimicrob. Agents Chemother. 55:983-991(2011) [PubMed] [Europe PMC] [Abstract]
Cited for: ENZYME REGULATION, LETHAL DOSE.
[10]"The crystal structure of Pseudomonas aeruginosa exotoxin domain III with nicotinamide and AMP: conformational differences with the intact exotoxin."
Li M., Dyda F., Benhar I., Pastan I., Davies D.R.
Proc. Natl. Acad. Sci. U.S.A. 92:9308-9312(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (3.0 ANGSTROMS) OF 424-638.
[11]"Crystal structure of the catalytic domain of Pseudomonas exotoxin A complexed with a nicotinamide adenine dinucleotide analog: implications for the activation process and for ADP ribosylation."
Li M., Dyda F., Benhar I., Pastan I., Davies D.R.
Proc. Natl. Acad. Sci. U.S.A. 93:6902-6906(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF 424-638.
[12]"The nature and character of the transition state for the ADP-ribosyltransferase reaction."
Jorgensen R., Wang Y., Visschedyk D., Merrill A.R.
EMBO Rep. 9:802-809(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.35 ANGSTROMS) OF 425-630 IN COMPLEX WITH YEAST EEF2 AND NAD, MUTAGENESIS OF ARG-481; GLU-571; ARG-576 AND GLU-578.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
K01397 Genomic DNA. Translation: AAB59097.1.
AE004091 Genomic DNA. Translation: AAG04537.1.
PIRA30347.
C83503.
RefSeqNP_249839.1. NC_002516.2.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1AERX-ray2.30A/B425-634[»]
1DMAX-ray2.50A/B425-638[»]
1IKPX-ray1.45A26-638[»]
1IKQX-ray1.62A26-638[»]
1XK9X-ray2.10A/B424-638[»]
1ZM2X-ray3.07B/D/F424-630[»]
1ZM3X-ray3.07B/D/F424-630[»]
1ZM4X-ray2.90B/D/F424-630[»]
1ZM9X-ray2.80B/D/F424-630[»]
2ZITX-ray3.00B/D/F425-630[»]
3B78X-ray2.50B/D/F425-630[»]
3B82X-ray2.35B/D/F425-630[»]
3B8HX-ray2.50B/D/F425-630[»]
ProteinModelPortalP11439.
SMRP11439. Positions 27-631.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

STRING208964.PA1148.

Chemistry

DrugBankDB00131. Adenosine monophosphate.

Protein family/group databases

TCDB1.C.73.1.1. the pseudomonas exotoxin a (p-exoa) family.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

GeneID877850.
KEGGpae:PA1148.
PATRIC19836616. VBIPseAer58763_1194.

Organism-specific databases

PseudoCAPPA1148.

Phylogenomic databases

HOGENOMHOG000081585.
KOK11020.
OMAAHESNEM.
OrthoDBEOG6ZPSVM.
ProtClustDBCLSK545809.

Enzyme and pathway databases

BioCycMetaCyc:MONOMER-15587.
SABIO-RKP11439.

Family and domain databases

Gene3D2.60.120.200. 1 hit.
3.90.1350.10. 1 hit.
InterProIPR008985. ConA-like_lec_gl_sf.
IPR013320. ConA-like_subgrp.
IPR015185. Exotox-A_bind.
IPR015099. Exotox-A_cataly_dom.
IPR015186. Exotox-A_middle_dom.
[Graphical view]
PfamPF09101. Exotox-A_bind. 1 hit.
PF09009. Exotox-A_cataly. 1 hit.
PF09102. Exotox-A_target. 1 hit.
[Graphical view]
SUPFAMSSF49899. SSF49899. 1 hit.
SSF56864. SSF56864. 1 hit.
ProtoNetSearch...

Other

EvolutionaryTraceP11439.

Entry information

Entry nameTOXA_PSEAE
AccessionPrimary (citable) accession number: P11439
Secondary accession number(s): Q9I4I7
Entry history
Integrated into UniProtKB/Swiss-Prot: October 1, 1989
Last sequence update: January 11, 2001
Last modified: April 16, 2014
This is version 119 of the entry and version 2 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references