Reviewed, UniProtKB/Swiss-Prot P15917 (LEF_BACAN)

Names and origin

Protein names

Recommended name:
    Lethal factor
      Short name=LF
    EC=3.4.24.83
Alternative name(s):
    Anthrax lethal toxin endopeptidase component

Gene names

Name:	lef
Ordered Locus Names:	pXO1-107, BXA0172, GBAA_pXO1_0172

Encoded on

Plasmid pXO1

Organism

Bacillus anthracis [Complete proteome] [HAMAP]

Taxonomic identifier

1392 [NCBI]

Taxonomic lineage

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

Protein attributes

Sequence length	809 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. LF is the lethal factor that, when associated with PA, causes death. LF is not toxic by itself. It is a protease that cleaves the N-terminal of most dual specificity mitogen-activated protein kinase kinases (MAPKKs or MAP2Ks) (except for MAP2K5). Cleavage invariably occurs within the N-terminal proline-rich region preceding the kinase domain, thus disrupting a sequence involved in directing specific protein-protein interactions necessary for the assembly of signaling complexes. There may be other cytosolic targets of LF involved in cytotoxicity. The proteasome may mediate a toxic process initiated by LF in the cell cytosol involving degradation of unidentified molecules that are essential for macrophage homeostasis. This is an early step in LeTx intoxication, but it is downstream of the cleavage by LF of MEK1 or other putative substrates. Ref.7 Ref.8 Ref.9 Ref.10 Ref.11
Catalytic activity	Preferred amino acids around the cleavage site can be denoted BBBBxHx-|-H, in which B denotes Arg or Lys, H denotes a hydrophobic amino acid, and x is any amino acid. The only known protein substrates are mitogen-activated protein (MAP) kinase kinases.
Cofactor	Binds 1 zinc ion per subunit.
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).
Subcellular location	Secreted.
Induction	Positively transcriptionally regulated by AtxA, which, in turn, is induced by bicarbonate and high temperatures (37 degrees Celsius).
Domain	It comprises four domains: domain I binds the membrane-translocating component (PA); domains II, III and IV together create a long deep groove that holds the 16-residue N-terminal tail of MAPKK before cleavage. Domain IV contains the catalytic center. The PA-binding region is found in both B.anthracis EF and LF.
Miscellaneous	LF 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. LeTx requires passage through an acidic vesicle for activity; at acidic pH, as the pore is inserted into the membrane, LF is translocated and reaches its cytosolic targets. LF is probably directly involved in its routing, by interacting with the lipid membrane. This interaction could involve a conformational change of LF and/or an oligomerization of the protein. LF may have the capability of partially unfolding in order to cross the membrane.
Sequence similarities	Belongs to the peptidase M34 family.

Ontologies

Keywords
Biological process	Virulence
Cellular component	Secreted
Domain	Repeat Signal
Ligand	Metal-binding Zinc
Molecular function	Hydrolase Metalloprotease Protease Toxin
Technical term	3D-structure Complete proteome Direct protein sequencing Plasmid
Gene Ontology (GO)
Biological process	pathogenesis Inferred from electronic annotation. Source: UniProtKB-KW proteolysis Inferred from electronic annotation. Source: InterPro
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	metallopeptidase activity Inferred from electronic annotation. Source: UniProtKB-KW protein binding Inferred from electronic annotation. Source: InterPro zinc ion binding Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 33

33

Ref.1

Chain

34 – 809

776

Lethal factor

PRO_0000029233

Regions

Repeat

315 – 333

19

1

Repeat

342 – 357

16

2

Repeat

360 – 378

19

3

Repeat

380 – 397

18

4

Repeat

399 – 416

18

5

Region

34 – 293

260

PA-binding region Potential

Region

60 – 295

236

I; PA-binding region Potential

Region

296 – 330

35

IIA

Region

315 – 416

102

5 X approximate repeats

Region

336 – 416

81

III

Region

420 – 583

164

IIB

Region

585 – 809

225

IV

Sites

Active site

720

1

Metal binding

719

1

Zinc; catalytic

Metal binding

723

1

Zinc; catalytic

Metal binding

768

1

Zinc; catalytic

Natural variations

Natural variant

299

1

A → S in strain: Sterne.

Experimental info

Mutagenesis

180

1

V → A: No effect on PA-binding ability. Ref.16

Mutagenesis

181

1

Y → A: Loss of ability to bind to PA. Ref.16

Mutagenesis

182

1

Y → A: Loss of ability to bind to PA. Ref.16

Mutagenesis

183

1

E → A: No effect on PA-binding ability. Ref.16

Mutagenesis

184

1

I → A: Loss of ability to bind to PA. Ref.16

Mutagenesis

185

1

G → A: No effect on PA-binding ability. Ref.16

Mutagenesis

186

1

K → A: Loss of ability to bind to PA. Ref.16

Mutagenesis

220

1

D → A: Loss of ability to bind to PA and loss of toxicity. Ref.17

Mutagenesis

221

1

L → A: No effect on PA-binding ability and fully toxic. Ref.17

Mutagenesis

222

1

L → A: No effect on PA-binding ability and fully toxic. Ref.17

Mutagenesis

223

1

F → A: Loss of ability to bind to PA and non-toxic. Ref.17

Mutagenesis

719

1

H → A: Loss of activity and zinc binding. Ref.15

Mutagenesis

720

1

E → C or D: Loss of activity. No effect on zinc binding. Ref.15

Mutagenesis

723

1

H → A: Loss of activity and zinc binding. Ref.15

Secondary structure

1

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809

Helix Strand Turn

Details...

Turn

62 – 65

4

Helix

66 – 76

11

Beta strand

77 – 80

4

Helix

87 – 97

11

Helix

101 – 109

9

Beta strand

113 – 119

7

Helix

121 – 123

3

Helix

125 – 127

3

Helix

132 – 135

4

Beta strand

136 – 138

3

Beta strand

144 – 146

3

Helix

147 – 149

3

Beta strand

151 – 155

5

Beta strand

157 – 159

3

Beta strand

161 – 165

5

Turn

170 – 172

3

Helix

174 – 190

17

Helix

193 – 196

4

Helix

201 – 211

11

Beta strand

213 – 216

4

Helix

217 – 222

6

Helix

225 – 228

4

Helix

236 – 240

5

Helix

243 – 258

16

Helix

260 – 269

10

Helix

271 – 282

12

Helix

284 – 293

10

Helix

297 – 310

14

Helix

312 – 316

5

Helix

320 – 330

11

Helix

337 – 341

5

Turn

342 – 346

5

Helix

347 – 353

7

Helix

357 – 359

3

Beta strand

361 – 363

3

Helix

365 – 375

11

Helix

406 – 415

10

Helix

421 – 428

8

Helix

439 – 455

17

Beta strand

469 – 474

6

Helix

476 – 478

3

Helix

481 – 484

4

Helix

498 – 505

8

Beta strand

510 – 513

4

Beta strand

518 – 522

5

Beta strand

532 – 537

6

Beta strand

543 – 547

5

Turn

548 – 550

3

Beta strand

551 – 554

4

Beta strand

556 – 570

15

Beta strand

573 – 583

11

Helix

585 – 606

22

Beta strand

616 – 619

4

Helix

625 – 642

18

Helix

645 – 657

13

Beta strand

662 – 667

6

Helix

669 – 671

3

Helix

676 – 678

3

Helix

682 – 684

3

Beta strand

688 – 693

6

Turn

694 – 697

4

Beta strand

698 – 705

8

Helix

713 – 731

19

Helix

745 – 753

9

Helix

762 – 764

3

Helix

766 – 777

12

Helix

782 – 791

10

Helix

793 – 802

10

Sequences

	Sequence		Length	Mass (Da)
	P15917-1 [UniParc]. Last modified July 5, 2004. Version 2. Checksum: 2076B4D7277317EE	FASTA	809	93,770
	10 20 30 40 50 60 MNIKKEFIKV ISMSCLVTAI TLSGPVFIPL VQGAGGHGDV GMHVKEKEKN KDENKRKDEE 70 80 90 100 110 120 RNKTQEEHLK EIMKHIVKIE VKGEEAVKKE AAEKLLEKVP SDVLEMYKAI GGKIYIVDGD 130 140 150 160 170 180 ITKHISLEAL SEDKKKIKDI YGKDALLHEH YVYAKEGYEP VLVIQSSEDY VENTEKALNV 190 200 210 220 230 240 YYEIGKILSR DILSKINQPY QKFLDVLNTI KNASDSDGQD LLFTNQLKEH PTDFSVEFLE 250 260 270 280 290 300 QNSNEVQEVF AKAFAYYIEP QHRDVLQLYA PEAFNYMDKF NEQEINLSLE ELKDQRMLAR 310 320 330 340 350 360 YEKWEKIKQH YQHWSDSLSE EGRGLLKKLQ IPIEPKKDDI IHSLSQEEKE LLKRIQIDSS 370 380 390 400 410 420 DFLSTEEKEF LKKLQIDIRD SLSEEEKELL NRIQVDSSNP LSEKEKEFLK KLKLDIQPYD 430 440 450 460 470 480 INQRLQDTGG LIDSPSINLD VRKQYKRDIQ NIDALLHQSI GSTLYNKIYL YENMNINNLT 490 500 510 520 530 540 ATLGADLVDS TDNTKINRGI FNEFKKNFKY SISSNYMIVD INERPALDNE RLKWRIQLSP 550 560 570 580 590 600 DTRAGYLENG KLILQRNIGL EIKDVQIIKQ SEKEYIRIDA KVVPKSKIDT KIQEAQLNIN 610 620 630 640 650 660 QEWNKALGLP KYTKLITFNV HNRYASNIVE SAYLILNEWK NNIQSDLIKK VTNYLVDGNG 670 680 690 700 710 720 RFVFTDITLP NIAEQYTHQD EIYEQVHSKG LYVPESRSIL LHGPSKGVEL RNDSEGFIHE 730 740 750 760 770 780 FGHAVDDYAG YLLDKNQSDL VTNSKKFIDI FKEEGSNLTS YGRTNEAEFF AEAFRLMHST 790 800 DHAERLKVQK NAPKTFQFIN DQIKFIINS

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Nucleotide sequence and analysis of the lethal factor gene (lef) from Bacillus anthracis." Bragg T.S., Robertson D.L. Gene 81:45-54(1989) [PubMed: 2509294] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 34-49.
[2]	"A comparison of Bacillus anthracis sequences." Lowe J. Submitted (APR-1990) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[3]	"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: 10515943] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Sterne.
[4]	"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: 12004073] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Ames / isolate Florida / A2012.
[5]	"Bacillus anthracis comparative genomics." Ravel J., Rasko D.A., Shumway M.F., Jiang L., Cer R.Z., Federova N.B., Wilson M., Stanley S., Decker S., Read T.D., Salzberg S.L., Fraser C.M. Submitted (MAY-2004) to the EMBL/GenBank/DDBJ databases Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Ames ancestor.
[6]	"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: 12067380] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 29-809. Strain: Carbosap and Ferrara.
[7]	"Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor." Duesbery N.S., Webb C.P., Leppla S.H., Gordon V.M., Klimpel K.R., Copeland T.D., Ahn N.G., Oskarsson M.K., Fukasawa K., Paull K.D., Vande Woude G.F. Science 280:734-737(1998) [PubMed: 9563949] [Abstract] Cited for: FUNCTION.
[8]	"Anthrax lethal factor cleaves the N-terminus of MAPKKs and induces tyrosine/threonine phosphorylation of MAPKs in cultured macrophages." Vitale G., Pellizzari R., Recchi C., Napolitani G., Mock M., Montecucco C. Biochem. Biophys. Res. Commun. 248:706-711(1998) [PubMed: 9703991] [Abstract] Cited for: FUNCTION.
[9]	"Anthrax lethal factor causes proteolytic inactivation of mitogen-activated protein kinase kinase." Duesbery N.S., Vande Woude G.F. J. Appl. Microbiol. 87:289-293(1999) [PubMed: 10475971] [Abstract] Cited for: FUNCTION.
[10]	"Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor." Vitale G., Bernardi L., Napolitani G., Mock M., Montecucco C. Biochem. J. 352:739-745(2000) [PubMed: 11104681] [Abstract] Cited for: FUNCTION.
[11]	"Proteasome activity is required for anthrax lethal toxin to kill macrophages." Tang G., Leppla S.H. Infect. Immun. 67:3055-3060(1999) [PubMed: 10338520] [Abstract] Cited for: FUNCTION.
[12]	"Secondary structure of anthrax lethal toxin proteins and their interaction with large unilamellar vesicles: a Fourier-transform infrared spectroscopy approach." Wang X.-M., Mock M., Ruysschaert J.-M., Cabiaux V. Biochemistry 35:14939-14946(1996) [PubMed: 8942659] [Abstract] Cited for: CHARACTERIZATION.
[13]	"Zinc content of the Bacillus anthracis lethal factor." Kochi S.K., Schiavo G., Mock M., Montecucco C. FEMS Microbiol. Lett. 124:343-348(1994) [PubMed: 7851740] [Abstract] Cited for: ZINC-BINDING.
[14]	"The three Bacillus anthracis toxin genes are coordinately regulated by bicarbonate and temperature." Sirard J.-C., Mock M., Fouet A. J. Bacteriol. 176:5188-5192(1994) [PubMed: 8051039] [Abstract] Cited for: REGULATION OF TOXIN EXPRESSION. Strain: Sterne.
[15]	"Lethal factor active-site mutations affect catalytic activity in vitro." Hammond S.E., Hanna P.C. Infect. Immun. 66:2374-2378(1998) [PubMed: 9573135] [Abstract] Cited for: MUTAGENESIS OF HIS-719; GLU-720 AND HIS-723. Strain: Sterne.
[16]	"Involvement of residues 147VYYEIGK153 in binding of lethal factor to protective antigen of Bacillus anthracis." Gupta P., Singh A., Chauhan V., Bhatnagar R. Biochem. Biophys. Res. Commun. 280:158-163(2001) [PubMed: 11162493] [Abstract] Cited for: MUTAGENESIS OF VAL-180; TYR-181; TYR-182; GLU-183; ILE-184; GLY-185 AND LYS-186. Strain: Sterne.
[17]	"Asp 187 and Phe 190 residues in lethal factor are required for the expression of anthrax lethal toxin activity." Singh A., Chauhan V., Sodhi A., Bhatnagar R. FEMS Microbiol. Lett. 212:183-186(2002) [PubMed: 12113932] [Abstract] Cited for: MUTAGENESIS OF ASP-220; LEU-221; LEU-222 AND PHE-223. Strain: Sterne.
[18]	"Toxins of Bacillus anthracis." Brossier F., Mock M. Toxicon 39:1747-1755(2001) [PubMed: 11595637] [Abstract] Cited for: REVIEW.
[19]	"Crystal structure of the anthrax lethal factor." Pannifer A.D., Wong T.Y., Schwarzenbacher R., Renatus M., Petosa C., Bienkowska J., Lacy D.B., Collier R.J., Park S., Leppla S.H., Hanna P.C., Liddington R.C. Nature 414:229-233(2001) [PubMed: 11700563] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) IN COMPLEX WITH ZINC IONS AND MAP2K2.
[20]	"The structural basis for substrate and inhibitor selectivity of the anthrax lethal factor." Turk B.E., Wong T.Y., Schwarzenbacher R., Jarrell E.T., Leppla S.H., Collier R.J., Liddington R.C., Cantley L.C. Nat. Struct. Mol. Biol. 11:60-66(2004) [PubMed: 14718924] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (3.52 ANGSTROMS) OF 34-809 IN COMPLEX WITH ZINC IONS AND PEPTIDE SUBSTRATE ANALOG.
[21]	"Anthrax lethal factor inhibition." Shoop W.L., Xiong Y., Wiltsie J., Woods A., Guo J., Pivnichny J.V., Felcetto T., Michael B.F., Bansal A., Cummings R.T., Cunningham B.R., Friedlander A.M., Douglas C.M., Patel S.B., Wisniewski D., Scapin G., Salowe S.P., Zaller D.M. , Chapman K.T., Scolnick E.M., Schmatz D.M., Bartizal K., MacCoss M., Hermes J.D. Proc. Natl. Acad. Sci. U.S.A. 102:7958-7963(2005) [PubMed: 15911756] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF 297-809 IN COMPLEX WITH ZINC IONS AND PROTEASE INHIBITOR.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

M29081 Genomic DNA. Translation: AAA79216.1.
M30210 Genomic DNA. Translation: AAA22569.1.
AF065404 Genomic DNA. Translation: AAD32411.1.
AE011190 Genomic DNA. Translation: AAM26117.1.
AE017336 Genomic DNA. Translation: AAT28913.2.
AJ413934 Genomic DNA. Translation: CAC93932.1.
AJ413935 Genomic DNA. Translation: CAC93933.1.

PIR

JQ0032.

RefSeq

NP_052803.1.
NP_652928.1.
YP_016503.2.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1J7N	X-ray	2.30	A/B	34-809	[»]
1JKY	X-ray	3.90	A	34-809	[»]
1PWP	X-ray	2.90	A/B	34-809	[»]
1PWQ	X-ray	3.52	A/B	34-809	[»]
1PWU	X-ray	2.70	A/B	34-809	[»]
1PWV	X-ray	2.85	A/B	34-809	[»]
1PWW	X-ray	2.80	A/B	34-809	[»]
1YQY	X-ray	2.30	A	297-809	[»]
1ZXV	X-ray	2.67	A/B	34-809	[»]

ModBase

Search...

Protein family/group databases

MEROPS

M34.001.

Genome annotation databases

GeneID

1158731.
2820148.
3361711.

GenomeReviews

Gene locus GBAA_pXO1_0172 in contig AE017336_GR.

KEGG

bar:GBAA_pXO1_0172.

TIGR

GBAA_pXO1_0172.

Enzyme and pathway databases

BioCyc

BANT261594:GBAA_PXO1_0162-MON.

BRENDA

3.4.24.83. 267517.

Pathway_Interaction_DB

anthraxpathway. Cellular roles of Anthrax toxin.

Family and domain databases

InterPro

IPR015239. Anthrax_tox_lethal-fac_cen.
IPR003541. Anthrax_toxin_lethal/edema_N.
IPR014781. Anthrax_toxin_lethal/edema_N/C.
IPR006025. Pept_M_Zn_BS.
[Graphical view]

Pfam

PF09156. Anthrax-tox_M. 1 hit.
PF07737. ATLF. 2 hits.
[Graphical view]

PRINTS

PR01392. ANTHRAXTOXNA.

PROSITE

PS00142. ZINC_PROTEASE. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

LEF_BACAN

Accession

Primary (citable) accession number: P15917
Secondary accession number(s): Q8KYJ6, Q933F6

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 1, 1990
Last sequence update:	July 5, 2004
Last modified:	July 28, 2009
This is version 103 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)