Calmodulin-sensitive adenylate cyclase precursor

Preferred order of sections

Names and origin

Protein names

Recommended name:
Calmodulin-sensitive adenylate cyclase
EC=4.6.1.1

Alternative name(s):
ATP pyrophosphate-lyase
Adenylyl cyclase
Anthrax edema toxin adenylate cyclase component
Edema factor
Short name=EF

Gene names

Name:	cya
Ordered Locus Names:	pXO1-122, BXA0141, GBAA_pXO1_0142

Encoded on

Plasmid pXO1

Organism

Bacillus anthracis [Reference proteome] [HAMAP]

Taxonomic identifier

1392 [NCBI]

Taxonomic lineage

Bacteria › Firmicutes › Bacilli › Bacillales › Bacillaceae › Bacillus › Bacillus cereus group

Protein attributes

Sequence length	800 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	One of the three proteins composing the anthrax toxin, the agent which infects many mammalian species and that may cause death. EF is a calmodulin-dependent adenylyl cyclase that, when associated with PA, causes edema. EF is not toxic by itself and it is required for the survival of germinated spores within macrophages at the early stages of infection. Provokes dramatic elevation of intracellular cAMP levels in the host.
Catalytic activity	ATP = 3',5'-cyclic AMP + diphosphate.
Cofactor	Calcium.
Enzyme regulation	It has an absolute requirement for host calmodulin for its activation. Inhibited by ethyl 5-aminopyrazolo[1,5-a]quinazoline-3-carboxylate.
Subunit structure	Anthrax toxins are composed of three distinct proteins, a protective antigen (PA), a lethal factor (LF) and an edema factor (EF). None of these is toxic by itself. PA+LF forms the lethal toxin (LeTx); PA+EF forms the edema toxin (EdTx). EF probably forms oligomers as part of the translocation machinery, formed by a heterocomplex of PA63 monomers and EF subunits, and it is functional as a monomer in the host cell.
Subcellular location	Secreted.
Domain	The N-terminal region contains the residues responsible for binding to PA63. The C-terminal region contains the calmodulin-dependent activation domain and the catalytic site. This region is composed of three globular domains: CA, CB and a helical domain connected to CA by a linker. The active site lies at the interface of CA and CB. The metal ion is coordinated by residues from CA; calmodulin probably binds in a multistep fashion first to residues in CA and then to residues present in the linker and the helical domain. Ref.11 The PA-binding region is found in both B.anthracis EF and LF. Ref.11
Miscellaneous	EF binds to the heptamer formed by cleaved PA on the host cell membrane. This step is followed by internalization of the hetero-oligomeric complex by receptor-mediated endocytosis. EF requires passage through an acidic vesicle for activity. At acidic pH, the pore is inserted into the membrane, allowing translocation of EF, which probably contributes actively to its own insertion into the membrane. EF remains associated to the vesicle membrane after translocation to the cytosol, with the catalytic domains being exposed on the cytoplasmic face.
Sequence similarities	Belongs to the adenylyl cyclase class-2 family.

Ontologies

Keywords
Biological process	Virulence cAMP biosynthesis
Cellular component	Secreted
Domain	Signal
Ligand	ATP-binding Calcium Calmodulin-binding Magnesium Metal-binding Nucleotide-binding
Molecular function	Lyase Toxin
Technical term	3D-structure Complete proteome Direct protein sequencing Plasmid Reference proteome
Gene Ontology (GO)
Biological_process	pathogenesis Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW calcium- and calmodulin-responsive adenylate cyclase activity Inferred from electronic annotation. Source: InterPro metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW metallopeptidase activity Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
calm2	P62155	2	EBI-457011,EBI-397568	From a different organism.
CALM3	P62158	8	EBI-457011,EBI-397435	From a different organism.

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 33

33

Ref.2

Chain

34 – 800

767

Calmodulin-sensitive adenylate cyclase

PRO_0000001317

Regions

Region

34 – 290

257

Interaction with protective antigen

Region

294 – 349

56

Catalytic CA1

Region

350 – 489

140

Catalytic CB

Region

490 – 622

133

Catalytic CA2

Region

623 – 800

178

Interaction with calmodulin

Sites

Active site

351

1

Proton acceptor

Metal binding

491

1

Magnesium

Metal binding

493

1

Magnesium

Experimental info

Mutagenesis

169

1

V → A: No effect. Ref.16

Mutagenesis

170

1

Y → A: Loss of cytotoxicity due to inability to bind PA. Ref.16

Mutagenesis

171

1

Y → A: Loss of cytotoxicity due to inability to bind PA. Ref.16

Mutagenesis

172

1

E → A: No effect. Ref.16

Mutagenesis

173

1

I → A: Loss of cytotoxicity due to inability to bind PA. Ref.16

Mutagenesis

174

1

G → A: No effect. Ref.16

Mutagenesis

175

1

K → A: Loss of cytotoxicity due to inability to bind PA. Ref.16

Mutagenesis

329

1

R → M: Great decrease in activity. Ref.17

Mutagenesis

346

1

K → M or R: Loss of activity. Ref.14 Ref.15

Mutagenesis

346

1

K → Q: Loss of activity due to inability to bind the substrate. Ref.14 Ref.15

Mutagenesis

353

1

K → M, R or A: Loss of activity. Ref.14

Mutagenesis

436

1

E → Q: Decreases activity. Ref.14

Mutagenesis

443

1

E → Q: Decreases activity. Ref.17

Mutagenesis

491

1

D → N: Great decrease in activity. Ref.17

Mutagenesis

493

1

D → N: Great decrease in activity. Ref.17

Mutagenesis

523

1

L → A: Little effect on activation by calmodulin.

Mutagenesis

525

1

K → A: Great decrease in calmodulin binding.

Mutagenesis

526

1

Q → A: Little effect on activation by calmodulin.

Mutagenesis

529

1

V → A: Little effect on activation by calmodulin.

Mutagenesis

577

1

H → N or D: Loss of function.

Mutagenesis

583

1

N → A: Decreases activity.

Mutagenesis

583

1

N → Q or H: Loss of function.

Mutagenesis

588

1

E → A: Loss of function.

Mutagenesis

590

1

D → A: Decreases activity.

Mutagenesis

639

1

N → A: Decreases catalysis rate.

Mutagenesis

647

1

D → A: Decreases activity due to reduced activation by calmodulin.

Sequence conflict

350

1

V → E in AAA79215. Ref.2

Sequence conflict

510

1

Q → T AA sequence Ref.2

Sequence conflict

512 – 513

2

EW → RM AA sequence Ref.2

Sequence conflict

760

1

V → L in CAA00652. Ref.3

Secondary structure

1

.

800

Helix Strand Turn

Details...

Sequences

Sequence

Length

Mass (Da)

Tools

P40136 [UniParc].

Last modified February 1, 1995. Version 1.
Checksum: F4F7EB485DF4C5A6
Show »

FASTA

800

92,478

        10         20         30         40         50         60 
MTRNKFIPNK FSIISFSVLL FAISSSQAIE VNAMNEHYTE SDIKRNHKTE KNKTEKEKFK 

        70         80         90        100        110        120 
DSINNLVKTE FTNETLDKIQ QTQDLLKKIP KDVLEIYSEL GGEIYFTDID LVEHKELQDL 

       130        140        150        160        170        180 
SEEEKNSMNS RGEKVPFASR FVFEKKRETP KLIINIKDYA INSEQSKEVY YEIGKGISLD 

       190        200        210        220        230        240 
IISKDKSLDP EFLNLIKSLS DDSDSSDLLF SQKFKEKLEL NNKSIDINFI KENLTEFQHA 

       250        260        270        280        290        300 
FSLAFSYYFA PDHRTVLELY APDMFEYMNK LEKGGFEKIS ESLKKEGVEK DRIDVLKGEK 

       310        320        330        340        350        360 
ALKASGLVPE HADAFKKIAR ELNTYILFRP VNKLATNLIK SGVATKGLNV HGKSSDWGPV 

       370        380        390        400        410        420 
AGYIPFDQDL SKKHGQQLAV EKGNLENKKS ITEHEGEIGK IPLKLDHLRI EELKENGIIL 

       430        440        450        460        470        480 
KGKKEIDNGK KYYLLESNNQ VYEFRISDEN NEVQYKTKEG KITVLGEKFN WRNIEVMAKN 

       490        500        510        520        530        540 
VEGVLKPLTA DYDLFALAPS LTEIKKQIPQ KEWDKVVNTP NSLEKQKGVT NLLIKYGIER 

       550        560        570        580        590        600 
KPDSTKGTLS NWQKQMLDRL NEAVKYTGYT GGDVVNHGTE QDNEEFPEKD NEIFIINPEG 

       610        620        630        640        650        660 
EFILTKNWEM TGRFIEKNIT GKDYLYYFNR SYNKIAPGNK AYIEWTDPIT KAKINTIPTS 

       670        680        690        700        710        720 
AEFIKNLSSI RRSSNVGVYK DSGDKDEFAK KESVKKIAGY LSDYYNSANH IFSQEKKRKI 

       730        740        750        760        770        780 
SIFRGIQAYN EIENVLKSKQ IAPEYKNYFQ YLKERITNQV QLLLTHQKSN IEFKLLYKQL 

       790        800 
NFTENETDNF EVFQKIIDEK

« Hide

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Structural homology between virulence-associated bacterial adenylate cyclases." Escuyer V., Duflot E., Sezer O., Danchin A., Mock M. Gene 71:293-298(1988) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"Nucleotide sequence of the Bacillus anthracis edema factor gene (cya): a calmodulin-dependent adenylate cyclase." Robertson D.L., Tippetts M.T., Leppla S.H. Gene 73:363-371(1988) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 34-48.
[3]	"Nucleotide sequences expressing adenylate cyclase from B.anthracis, proteins having the activity of this adenylate cyclase and biological uses." Escuyer V., Duflot E., Mock M., Danchin A. Patent number EP0366550, 02-MAY-1990 Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[4]	"Molecular cloning and expression of the Bacillus anthracis edema factor toxin gene: a calmodulin-dependent adenylate cyclase." Tippetts M.T., Robertson D.L. J. Bacteriol. 170:2263-2266(1988) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[5]	"Sequence and organization of pXO1, the large Bacillus anthracis plasmid harboring the anthrax toxin genes." Okinaka R.T., Cloud K., Hampton O., Hoffmaster A.R., Hill K.K., Keim P., Koehler T.M., Lamke G., Kumano S., Mahillon J., Manter D., Martinez Y., Ricke D., Svensson R., Jackson P.J. J. Bacteriol. 181:6509-6515(1999) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Sterne.
[6]	"Comparative genome sequencing for discovery of novel polymorphisms in Bacillus anthracis." Read T.D., Salzberg S.L., Pop M., Shumway M.F., Umayam L., Jiang L., Holtzapple E., Busch J.D., Smith K.L., Schupp J.M., Solomon D., Keim P., Fraser C.M. Science 296:2028-2033(2002) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Ames / isolate Florida / A2012.
[7]	"The complete genome sequence of Bacillus anthracis Ames 'Ancestor'." Ravel J., Jiang L., Stanley S.T., Wilson M.R., Decker R.S., Read T.D., Worsham P., Keim P.S., Salzberg S.L., Fraser-Liggett C.M., Rasko D.A. J. Bacteriol. 191:445-446(2009) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Ames ancestor.
[8]	"Sequence analysis of the genes encoding for the major virulence factors of Bacillus anthracis vaccine strain 'Carbosap'." Adone R., Pasquali P., La Rosa G., Marianelli C., Muscillo M., Fasanella A., Francia M., Ciuchini F. J. Appl. Microbiol. 93:117-121(2002) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-783. Strain: Carbosap and Ferrara.
[9]	"Structure and interaction of PA63 and EF (edema toxin) of Bacillus anthracis with lipid membrane." Wang X.-M., Wattiez R., Mock M., Falmagne P., Ruysschaert J.-M., Cabiaux V. Biochemistry 36:14906-14913(1997) [PubMed] [Europe PMC] [Abstract] Cited for: CHARACTERIZATION. Strain: Sterne.
[10]	"Translocation of Bacillus anthracis lethal and oedema factors across endosome membranes." Guidi-Rontani C., Weber-Levy M., Mock M., Cabiaux V. Cell. Microbiol. 2:259-264(2000) [PubMed] [Europe PMC] [Abstract] Cited for: CHARACTERIZATION. Strain: Sterne.
[11]	"Characterization of ATP and calmodulin-binding properties of a truncated form of Bacillus anthracis adenylate cyclase." Labruyere E., Mock M., Ladant D., Michelson S., Gilles A.-M., Laoide B., Barzu O. Biochemistry 29:4922-4928(1990) [PubMed] [Europe PMC] [Abstract] Cited for: CHARACTERIZATION OF CALMODULIN-BINDING DOMAIN.
[12]	"Fate of germinated Bacillus anthracis spores in primary murine macrophages." Guidi-Rontani C., Levy M., Ohayon H., Mock M. Mol. Microbiol. 42:931-938(2001) [PubMed] [Europe PMC] [Abstract] Cited for: EFFECT ON SPORE GERMINATION. Strain: Sterne.
[13]	"Structure-based inhibitor discovery against adenylyl cyclase toxins from pathogenic bacteria that cause anthrax and whooping cough." Soelaiman S., Wei B.Q., Bergson P., Lee Y.-S., Shen Y., Mrksich M., Shoichet B.K., Tang W.-J. J. Biol. Chem. 278:25990-25997(2003) [PubMed] [Europe PMC] [Abstract] Cited for: INHIBITION BY QUINAZOLINE COMPOUNDS.
[14]	"A-type ATP binding consensus sequences are critical for the catalytic activity of the calmodulin-sensitive adenylyl cyclase from Bacillus anthracis." Xia Z., Storm D.R. J. Biol. Chem. 265:6517-6520(1990) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF LYS-346; LYS-353 AND GLU-436. Strain: SRI-1.
[15]	"Structural and ligand-binding properties of a truncated form of Bacillus anthracis adenylate cyclase and of a catalytically inactive variant in which glutamine substitutes for lysine-346." Labruyere E., Mock M., Surewicz W.K., Mantsch H.H., Rose T., Munier H., Sarfati R.S., Barzu O. Biochemistry 30:2619-2624(1991) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF LYS-346. Strain: Sterne.
[16]	"Purification of anthrax edema factor from Escherichia coli and identification of residues required for binding to anthrax protective antigen." Kumar P., Ahuja N., Bhatnagar R. Infect. Immun. 69:6532-6536(2001) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF VAL-169; TYR-170; TYR-171; GLU-172; ILE-173; GLY-174 AND LYS-175.
[17]	"An extended conformation of calmodulin induces interactions between the structural domains of adenylyl cyclase from Bacillus anthracis to promote catalysis." Drum C.L., Yan S.-Z., Sarac R., Mabuchi Y., Beckingham K., Bohm A., Grabarek Z., Tang W.-J. J. Biol. Chem. 275:36334-36340(2000) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF ARG-329; GLU-443; ASP-491 AND ASP-493.
[18]	"Structural basis for the activation of anthrax adenylyl cyclase exotoxin by calmodulin." Drum C.L., Yan S.-Z., Bard J., Shen Y.-Q., Lu D., Soelaiman S., Grabarek Z., Bohm A., Tang W.-J. Nature 415:396-402(2002) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.6 ANGSTROMS), MUTAGENESIS.
[19]	"Phylogeny of adenylyl cyclases." Danchin A. Adv. Second Messenger Phosphoprotein Res. 27:109-162(1993) [PubMed] [Europe PMC] [Abstract] Cited for: REVIEW.
[20]	"Anthrax." Mock M., Fouet A. Annu. Rev. Microbiol. 55:647-671(2001) [PubMed] [Europe PMC] [Abstract] Cited for: REVIEW.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M23179 Genomic DNA. Translation: AAA22374.1.
M24074 Genomic DNA. Translation: AAA79215.1.
A07289 Unassigned DNA. Translation: CAA00652.1. Sequence problems.
AF065404 Genomic DNA. Translation: AAD32426.1.
AE011190 Genomic DNA. Translation: AAM26089.1.
AE017336 Genomic DNA. Translation: AAT28883.2.
AJ413930 Genomic DNA. Translation: CAC93924.1.
AJ413931 Genomic DNA. Translation: CAC93925.1.

PIR

B59106.
JS0029.

RefSeq

NP_052818.1. NC_001496.1.
NP_652900.1. NC_003980.1.
YP_016473.2. NC_007322.2.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1K8T	X-ray	2.60	A	291-800	[»]
1K90	X-ray	2.75	A/B/C	291-800	[»]
1K93	X-ray	2.95	A/B/C	291-800	[»]
1LVC	X-ray	3.60	A/B/C	291-800	[»]
1PK0	X-ray	3.30	A/B/C	292-798	[»]
1S26	X-ray	3.00	A/B/C	291-800	[»]
1SK6	X-ray	3.20	A/B/C	291-800	[»]
1XFU	X-ray	3.35	A/B/C/D/E/F	64-800	[»]
1XFV	X-ray	3.35	A/B/C/D/E/F	33-800	[»]
1XFW	X-ray	3.40	A/B/C/D/E/F	33-800	[»]
1XFX	X-ray	3.20	A/B/C/D/E/F	33-800	[»]
1XFY	X-ray	3.30	A/B/C/D/E/F	33-800	[»]
1XFZ	X-ray	3.25	A/B/C/D/E/F	33-800	[»]
1Y0V	X-ray	3.60	A/B/C/D/E/F	33-800	[»]

DisProt

DP00395.

ProteinModelPortal

P40136.

SMR

P40136. Positions 64-798.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-31055N.

IntAct

P40136. 3 interactions.

STRING

261594.GBAA_pXO1_0142.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAM26089; AAM26089; BX_A0142.
AAT28883; AAT28883; GBAA_pXO1_0142.

GeneID

1158701.
2820138.
3361726.

KEGG

bar:GBAA_pXO1_0142.

PATRIC

24662079. VBIBacAnt106580_0129.

Phylogenomic databases

eggNOG

NOG146287.

HOGENOM

HOG000034573.

KO

K11029.

OMA

QYLKEMI.

ProtClustDB

CLSK687645.

Enzyme and pathway databases

BioCyc

BANT261594:GJ7F-5728-MONOMER.

Pathway_Interaction_DB

anthraxpathway. Cellular roles of Anthrax toxin.

Family and domain databases

Gene3D

3.40.390.10. 1 hit.

InterPro

IPR005165. Anthrax_toxin_edema_cen.
IPR003541. Anthrax_toxin_lethal/edema.
IPR014781. Anthrax_toxin_lethal/edema_N/C.
IPR024079. MetalloPept_cat_dom.
[Graphical view]

Pfam

PF03497. Anthrax_toxA. 1 hit.
PF07737. ATLF. 1 hit.
[Graphical view]

PRINTS

PR01392. ANTHRAXTOXNA.

ProtoNet

Search...

Other

BindingDB

P40136.

ChEMBL

CHEMBL5396.

EvolutionaryTrace

P40136.

Entry information

Entry name

CYAA_BACAN

Accession

Primary (citable) accession number: P40136
Secondary accession number(s): Q937W4, Q937W5

Entry history

Integrated into UniProtKB/Swiss-Prot:	February 1, 1995
Last sequence update:	February 1, 1995
Last modified:	May 1, 2013
This is version 125 of the entry and version 1 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

SIMILARITY comments

Index of protein domains and families

Preferred order of sections

Names and origin

Protein attributes

General annotation (Comments)

Ontologies

Binary interactions

With

Entry

#Exp.

IntAct

Notes

Sequence annotation (Features)

Molecule processing

Regions

Sites

Experimental info

Secondary structure

Sequences

References

Cross-references

Sequence databases

3D structure databases

Protein-protein interaction databases

Protocols and materials databases

Genome annotation databases

Phylogenomic databases

Enzyme and pathway databases

Family and domain databases

Other

Entry information

Relevant documents