ID   HLA_STAAU               Reviewed;         319 AA.
AC   P09616;
DT   01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT   01-DEC-1992, sequence version 2.
DT   24-JAN-2024, entry version 150.
DE   RecName: Full=Alpha-hemolysin;
DE            Short=Alpha-HL;
DE   AltName: Full=Alpha-toxin;
DE   Flags: Precursor;
GN   Name=hly; Synonyms=hla;
OS   Staphylococcus aureus.
OC   Bacteria; Bacillota; Bacilli; Bacillales; Staphylococcaceae;
OC   Staphylococcus.
OX   NCBI_TaxID=1280;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 27-44.
RC   STRAIN=ATCC 10832 / Wood 46;
RX   PubMed=6500704; DOI=10.1128/iai.46.2.615-618.1984;
RA   Gray G.S., Kehoe M.;
RT   "Primary sequence of the alpha-toxin gene from Staphylococcus aureus wood
RT   46.";
RL   Infect. Immun. 46:615-618(1984).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND SEQUENCE REVISION.
RC   STRAIN=ATCC 10832 / Wood 46;
RA   Hedengrahn G.;
RL   Submitted (OCT-1992) to the EMBL/GenBank/DDBJ databases.
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 27-319, FUNCTION, SUBUNIT, AND
RP   MUTAGENESIS OF 315-GLU--ASN-319.
RC   STRAIN=ATCC 10832 / Wood 46;
RX   PubMed=1587866; DOI=10.1016/s0021-9258(19)50103-x;
RA   Walker B., Krishnasastry M., Zorn L., Kasianowicz J., Bayley H.;
RT   "Functional expression of the alpha-hemolysin of Staphylococcus aureus in
RT   intact Escherichia coli and in cell lysates. Deletion of five C-terminal
RT   amino acids selectively impairs hemolytic activity.";
RL   J. Biol. Chem. 267:10902-10909(1992).
RN   [4] {ECO:0007744|PDB:7AHL}
RP   X-RAY CRYSTALLOGRAPHY (1.89 ANGSTROMS) OF 27-319, SUBUNIT, SUBCELLULAR
RP   LOCATION, AND DOMAIN.
RC   STRAIN=ATCC 10832 / Wood 46;
RX   PubMed=8943190; DOI=10.1126/science.274.5294.1859;
RA   Song L., Hobaugh M.R., Shustak C., Cheley S., Bayley H., Gouaux J.E.;
RT   "Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane
RT   pore.";
RL   Science 274:1859-1866(1996).
RN   [5]
RP   SUBUNIT, AND MUTAGENESIS OF 28-ASP-SER-29; 28-ASP--ASP-39; 28-ASP--GLY-49;
RP   28-ASP--PHE-65; 312-TRP--ASN-319; 315-GLU--ASN-319 AND 317-MET--ASN-319.
RX   PubMed=1400487; DOI=10.1016/s0021-9258(19)36680-3;
RA   Walker B., Krishnasastry M., Zorn L., Bayley H.;
RT   "Assembly of the oligomeric membrane pore formed by Staphylococcal alpha-
RT   hemolysin examined by truncation mutagenesis.";
RL   J. Biol. Chem. 267:21782-21786(1992).
RN   [6]
RP   MUTAGENESIS OF HIS-61; HIS-74; HIS-170 AND HIS-285.
RC   STRAIN=8325-4;
RX   PubMed=8168947; DOI=10.1128/iai.62.5.1843-1847.1994;
RA   Menzies B.E., Kernodle D.S.;
RT   "Site-directed mutagenesis of the alpha-toxin gene of Staphylococcus
RT   aureus: role of histidines in toxin activity in vitro and in a murine
RT   model.";
RL   Infect. Immun. 62:1843-1847(1994).
RN   [7]
RP   MUTAGENESIS OF HIS-61; HIS-74; HIS-170 AND HIS-285.
RX   PubMed=8188346; DOI=10.1128/iai.62.6.2249-2256.1994;
RA   Jursch R., Hildebrand A., Hobom G., Tranum-Jensen J., Ward R., Kehoe M.,
RA   Bhakdi S.;
RT   "Histidine residues near the N-terminus of staphylococcal alpha-toxin as
RT   reporters of regions that are critical for oligomerization and pore
RT   formation.";
RL   Infect. Immun. 62:2249-2256(1994).
RN   [8]
RP   MUTAGENESIS.
RX   PubMed=7559447; DOI=10.1074/jbc.270.39.23065;
RA   Walker B., Bayley H.;
RT   "Key residues for membrane binding, oligomerization, and pore forming
RT   activity of staphylococcal alpha-hemolysin identified by cysteine scanning
RT   mutagenesis and targeted chemical modification.";
RL   J. Biol. Chem. 270:23065-23071(1995).
RN   [9]
RP   FUNCTION, INTERACTION WITH HUMAN ADAM10, AND MUTAGENESIS OF HIS-61.
RX   PubMed=20624979; DOI=10.1073/pnas.1001815107;
RA   Wilke G.A., Bubeck Wardenburg J.;
RT   "Role of a disintegrin and metalloprotease 10 in Staphylococcus aureus
RT   alpha-hemolysin-mediated cellular injury.";
RL   Proc. Natl. Acad. Sci. U.S.A. 107:13473-13478(2010).
CC   -!- FUNCTION: Alpha-toxin binds to the membrane of eukaryotic cells
CC       (particularly red blood cells, RBC) forming pores, resulting in
CC       hemolysis, with the release of low-molecular weight molecules leading
CC       to eventual osmotic RBC lysis (PubMed:1587866, PubMed:20624979). Human
CC       RBCs bind much less alpha-toxin than do rabbit RBCs (PubMed:1587866,
CC       PubMed:20624979). Heptamer oligomerization and pore formation is
CC       required for lytic activity (PubMed:1587866, PubMed:20624979).
CC       {ECO:0000269|PubMed:1587866, ECO:0000269|PubMed:20624979}.
CC   -!- SUBUNIT: Self-assembles to first form a non-lytic oligomeric
CC       intermediate, and then, a mushroom-shaped homoheptamer structure of 100
CC       Angstroms in length and up to 100 Angstroms in diameter
CC       (PubMed:8943190) (Probable). Interacts with human ADAM10; this
CC       interaction is required for toxin pore formation, disruption of focal
CC       adhesions, and hly-mediated cytotoxicity (PubMed:20624979).
CC       {ECO:0000269|PubMed:20624979, ECO:0000269|PubMed:8943190,
CC       ECO:0000305|PubMed:1400487, ECO:0000305|PubMed:1587866}.
CC   -!- INTERACTION:
CC       P09616; P09616: hly; NbExp=6; IntAct=EBI-15848589, EBI-15848589;
CC   -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:8943190}.
CC       Note=Secreted as a monomer (PubMed:8943190). After oligomerization and
CC       pore formation, the complex is translocated across the bilayer,
CC       probably via the Gly-rich domain of each strand.
CC       {ECO:0000269|PubMed:8943190}.
CC   -!- DOMAIN: The mushroom-shaped heptamer is composed of a cap domain
CC       (comprising 7 beta sandwiches and the amino latches of each protomer),
CC       7 rim regions whose protruding strands may interact with the membrane
CC       bilayer, and the stem domain (52 Angstroms in length, 26 Angstroms in
CC       diameter) which forms the transmembrane pore.
CC       {ECO:0000269|PubMed:8943190}.
CC   -!- SIMILARITY: Belongs to the aerolysin family. {ECO:0000305}.
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DR   EMBL; X01645; CAA25801.1; -; Genomic_DNA.
DR   EMBL; M90536; AAA26598.1; -; Genomic_DNA.
DR   PIR; S69209; S69209.
DR   PDB; 3M2L; X-ray; 2.10 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 3M3R; X-ray; 2.20 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 3M4D; X-ray; 1.90 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 3M4E; X-ray; 2.30 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 4IDJ; X-ray; 3.36 A; A=27-319.
DR   PDB; 4YHD; X-ray; 2.80 A; A/B/C/D/E/G=27-319.
DR   PDB; 6U3T; X-ray; 2.79 A; A/B=27-319.
DR   PDB; 6U49; X-ray; 2.35 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 6U4P; X-ray; 2.49 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 7AHL; X-ray; 1.89 A; A/B/C/D/E/F/G=27-319.
DR   PDB; 7O1Q; EM; 3.40 A; A/B/C/D/E/F/G=27-131, A/B/C/D/E/F/G=174-319.
DR   PDBsum; 3M2L; -.
DR   PDBsum; 3M3R; -.
DR   PDBsum; 3M4D; -.
DR   PDBsum; 3M4E; -.
DR   PDBsum; 4IDJ; -.
DR   PDBsum; 4YHD; -.
DR   PDBsum; 6U3T; -.
DR   PDBsum; 6U49; -.
DR   PDBsum; 6U4P; -.
DR   PDBsum; 7AHL; -.
DR   PDBsum; 7O1Q; -.
DR   AlphaFoldDB; P09616; -.
DR   EMDB; EMD-12696; -.
DR   SMR; P09616; -.
DR   DIP; DIP-59322N; -.
DR   IntAct; P09616; 1.
DR   ChEMBL; CHEMBL1075259; -.
DR   TCDB; 1.C.3.1.1; the Alpha-hemolysin channel-forming toxin (Alphahl) family.
DR   ABCD; P09616; 22 sequenced antibodies.
DR   Reactome; R-HSA-844456; The NLRP3 inflammasome.
DR   Reactome; R-HSA-9660826; Purinergic signaling in leishmaniasis infection.
DR   EvolutionaryTrace; P09616; -.
DR   PHI-base; PHI:10241; -.
DR   PHI-base; PHI:11285; -.
DR   PRO; PR:P09616; -.
DR   GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR   GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR   GO; GO:0090729; F:toxin activity; IMP:UniProtKB.
DR   GO; GO:0051715; P:cytolysis in another organism; IEA:InterPro.
DR   Gene3D; 2.70.240.10; Leukocidin/porin MspA; 1.
DR   InterPro; IPR005831; Aerolysin/haemolysin_CS.
DR   InterPro; IPR003963; Bi-component_toxin_staph.
DR   InterPro; IPR016183; Leukocidin/Hemolysin_toxin.
DR   InterPro; IPR036435; Leukocidin/porin_MspA_sf.
DR   NCBIfam; TIGR01002; hlyII; 1.
DR   Pfam; PF07968; Leukocidin; 1.
DR   PRINTS; PR01468; BICOMPNTOXIN.
DR   SUPFAM; SSF56959; Leukocidin-like; 1.
DR   PROSITE; PS00274; AEROLYSIN; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Cytolysis; Direct protein sequencing; Hemolysis; Secreted;
KW   Signal; Toxin; Virulence.
FT   SIGNAL          1..26
FT                   /evidence="ECO:0000269|PubMed:6500704"
FT   CHAIN           27..319
FT                   /note="Alpha-hemolysin"
FT                   /id="PRO_0000035636"
FT   MUTAGEN         28..65
FT                   /note="Missing: Binds host red blood cells, complete loss
FT                   of hemolytic activity, no longer oligomerizes."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   MUTAGEN         28..49
FT                   /note="Missing: Binds host red blood cells, complete loss
FT                   of hemolytic activity, forms hexamers."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   MUTAGEN         28..39
FT                   /note="Missing: Binds host red blood cells, complete loss
FT                   of hemolytic activity, mainly forms pentamers."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   MUTAGEN         28..29
FT                   /note="Missing: Binds host red blood cells, complete loss
FT                   of hemolytic activity, mainly forms pentamers."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   MUTAGEN         61
FT                   /note="H->I,L,P,R,S,T: No oligomerization nor hemolytic
FT                   activity, wild-type target cell-binding."
FT                   /evidence="ECO:0000269|PubMed:8188346"
FT   MUTAGEN         61
FT                   /note="H->L: No hemolytic activity, reduced
FT                   oligomerization, not toxic in mice, no effect on ADAM10
FT                   binding."
FT                   /evidence="ECO:0000269|PubMed:20624979,
FT                   ECO:0000269|PubMed:8168947"
FT   MUTAGEN         74
FT                   /note="H->I,R,T: No oligomerization, altered hemolysis,
FT                   near wild-type target cell binding."
FT                   /evidence="ECO:0000269|PubMed:8188346"
FT   MUTAGEN         74
FT                   /note="H->L: 7% of normal hemolytic activity, reduced
FT                   toxicity in mice."
FT                   /evidence="ECO:0000269|PubMed:8168947"
FT   MUTAGEN         170
FT                   /note="H->L,P,R: Decreased hemolysis, wild-type
FT                   oligomerization and target cell binding."
FT                   /evidence="ECO:0000269|PubMed:8188346"
FT   MUTAGEN         170
FT                   /note="H->L: 16% of normal hemolytic activity."
FT                   /evidence="ECO:0000269|PubMed:8168947"
FT   MUTAGEN         285
FT                   /note="H->I,P,R,S: Nearly wild-type oligomerization,
FT                   hemolysis and target cell binding."
FT                   /evidence="ECO:0000269|PubMed:8188346"
FT   MUTAGEN         285
FT                   /note="H->L: 46% of normal hemolytic activity, slowly toxic
FT                   in mice."
FT                   /evidence="ECO:0000269|PubMed:8168947"
FT   MUTAGEN         312..319
FT                   /note="Missing: Binds host red blood cells, complete loss
FT                   of hemolytic activity, no longer oligomerizes."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   MUTAGEN         315..319
FT                   /note="Missing: Binds host red blood cells, almost complete
FT                   loss of hemolytic activity, greatly reduced
FT                   oligomerization."
FT                   /evidence="ECO:0000269|PubMed:1400487,
FT                   ECO:0000269|PubMed:1587866"
FT   MUTAGEN         317..319
FT                   /note="Missing: Binds host red blood cells, almost complete
FT                   loss of hemolytic activity, greatly reduced
FT                   oligomerization."
FT                   /evidence="ECO:0000269|PubMed:1400487"
FT   HELIX           28..31
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   TURN            35..38
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          40..43
FT                   /evidence="ECO:0007829|PDB:4IDJ"
FT   STRAND          47..55
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   TURN            56..59
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          60..69
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          74..87
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          92..95
FT                   /evidence="ECO:0007829|PDB:3M4D"
FT   TURN            98..100
FT                   /evidence="ECO:0007829|PDB:6U3T"
FT   STRAND          101..115
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          123..129
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          135..153
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          158..184
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          188..197
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          200..202
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          205..208
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   TURN            215..217
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   HELIX           232..234
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   HELIX           239..241
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   HELIX           244..247
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          254..260
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          268..286
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          288..311
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   TURN            312..315
FT                   /evidence="ECO:0007829|PDB:7AHL"
FT   STRAND          316..318
FT                   /evidence="ECO:0007829|PDB:7AHL"
SQ   SEQUENCE   319 AA;  35904 MW;  6711C415DF7EBF30 CRC64;
     MKTRIVSSVT TTLLLGSILM NPVAGAADSD INIKTGTTDI GSNTTVKTGD LVTYDKENGM
     HKKVFYSFID DKNHNKKLLV IRTKGTIAGQ YRVYSEEGAN KSGLAWPSAF KVQLQLPDNE
     VAQISDYYPR NSIDTKEYMS TLTYGFNGNV TGDDTGKIGG LIGANVSIGH TLKYVQPDFK
     TILESPTDKK VGWKVIFNNM VNQNWGPYDR DSWNPVYGNQ LFMKTRNGSM KAADNFLDPN
     KASSLLSSGF SPDFATVITM DRKASKQQTN IDVIYERVRD DYQLHWTSTN WKGTNTKDKW
     TDRSSERYKI DWEKEEMTN
//