Exotoxin A precursor - Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)

Preferred order of sections

Names and origin

Protein names

Recommended 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

Organism

Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) [Reference proteome] [HAMAP]

Taxonomic identifier

208964 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Pseudomonadales › Pseudomonadaceae › Pseudomonas ›

Protein attributes

Sequence length	638 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence 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⁺ + peptide diphthamide = nicotinamide + peptide N-(ADP-D-ribosyl)diphthamide.
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. K_M=121 µM for NAD

Ontologies

Keywords
Domain	Signal
Ligand	NAD
Molecular function	Glycosyltransferase Toxin Transferase
PTM	Disulfide bond
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Molecular_function	NAD+-diphthamide ADP-ribosyltransferase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 25

25

Ref.1

Chain

26 – 638

613

Exotoxin A

PRO_0000019365

Regions

Nucleotide binding

465 – 467

3

NAD

Nucleotide binding

479 – 485

7

NAD

Region

26 – 277

252

IA (required for target cell recognition)

Region

278 – 389

112

II (required for translocation in target cell cytoplasm)

Region

390 – 429

40

IB

Region

430 – 638

209

III (required for ADP-ribosyl activity)

Sites

Active site

578

1

Ref.3

Binding site

474

1

NAD

Binding site

578

1

NAD

Amino acid modifications

Disulfide bond

290 ↔ 312

Ref.7

Experimental info

Mutagenesis

82

1

K → E: Loss of toxicity, no binding to LRP1 receptor. Ref.6

Mutagenesis

481

1

R → H: 52-fold decrease in ADPRT activity, 55-fold reduction in GH activity, 31-fold increase in dissociation constant for NAD(+). Ref.12

Mutagenesis

571

1

E → A: 833-fold decrease in ADPRT activity, 4-fold reduction in GH activity, 2-fold increase in dissociation constant for NAD(+). Ref.12

Mutagenesis

576

1

R → A: 2-fold decrease in ADPRT activity, 14-fold reduction in GH activity, 3-fold increase in dissociation constant for NAD(+). Ref.12

Mutagenesis

578

1

E → 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 conflict

4

1

T → I in AAB59097. Ref.1

Sequence conflict

22

1

F → S in AAB59097. Ref.1

Sequence conflict

204

1

A → T in AAB59097. Ref.1

Sequence conflict

389

1

S → N in AAB59097. Ref.1

Sequence conflict

432

1

I → V in AAB59097. Ref.1

Sequence conflict

540

1

G → S in AAB59097. Ref.1

Secondary structure

1

.

638

Helix Strand Turn

Details...

Sequences

Sequence

Length

Mass (Da)

Tools

P11439 [UniParc].

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

638

69,284

        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' referencesShow '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. 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.

PIR

A30347.
C83503.

RefSeq

NP_249839.1. NC_002516.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1AER	X-ray	2.30	A/B	425-634	[»]
1DMA	X-ray	2.50	A/B	425-638	[»]
1IKP	X-ray	1.45	A	26-638	[»]
1IKQ	X-ray	1.62	A	26-638	[»]
1XK9	X-ray	2.10	A/B	424-638	[»]
1ZM2	X-ray	3.07	B/D/F	424-630	[»]
1ZM3	X-ray	3.07	B/D/F	424-630	[»]
1ZM4	X-ray	2.90	B/D/F	424-630	[»]
1ZM9	X-ray	2.80	B/D/F	424-630	[»]
2ZIT	X-ray	3.00	B/D/F	425-630	[»]
3B78	X-ray	2.50	B/D/F	425-630	[»]
3B82	X-ray	2.35	B/D/F	425-630	[»]
3B8H	X-ray	2.50	B/D/F	425-630	[»]

ProteinModelPortal

P11439.

SMR

P11439. Positions 27-631.

ModBase

Search...

Protein-protein interaction databases

STRING

208964.PA1148.

Protein family/group databases

TCDB

1.C.73.1.1. pseudomonas exotoxin A (P-ExoA) family.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

GeneID

877850.

KEGG

pae:PA1148.

PATRIC

19836616. VBIPseAer58763_1194.

Organism-specific databases

PseudoCAP

PA1148.

Phylogenomic databases

HOGENOM

HOG000081585.

KO

K11020.

OMA

AHESNEM.

ProtClustDB

CLSK545809.

Enzyme and pathway databases

BioCyc

MetaCyc:MONOMER-15587.

SABIO-RK

P11439.

Family and domain databases

Gene3D

2.60.120.200. 1 hit.
3.90.1350.10. 1 hit.

InterPro

IPR008985. ConA-like_lec_gl_sf.
IPR013320. ConA-like_subgrp.
IPR015185. Exotox-A_bind.
IPR015099. Exotox-A_cataly.
IPR015186. Exotox-A_middle_dom.
[Graphical view]

Pfam

PF09101. Exotox-A_bind. 1 hit.
PF09009. Exotox-A_cataly. 1 hit.
PF09102. Exotox-A_target. 1 hit.
[Graphical view]

SUPFAM

SSF49899. ConA_like_lec_gl. 1 hit.
SSF56864. Exotox-A_target. 1 hit.

ProtoNet

Search...

Other

DrugBank

DB00131. Adenosine monophosphate.

EvolutionaryTrace

P11439.

Entry information

Entry name

TOXA_PSEAE

Accession

Primary (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:	May 1, 2013
This is version 112 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references