TXL_EISFE
ID TXL_EISFE Reviewed; 297 AA.
AC O18423;
DT 01-JUL-2008, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1998, sequence version 1.
DT 03-AUG-2022, entry version 70.
DE RecName: Full=Lysenin {ECO:0000303|PubMed:9210594};
DE AltName: Full=Beta-barrel pore-forming toxin {ECO:0000303|PubMed:27176125};
DE Short=Beta-PFT {ECO:0000303|PubMed:27048994, ECO:0000303|PubMed:27176125};
DE AltName: Full=Invertebrate cytolysin pore {ECO:0000303|PubMed:27176125};
DE AltName: Full=efL1 {ECO:0000303|PubMed:9210594};
OS Eisenia fetida (Red wiggler worm).
OC Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Annelida; Clitellata;
OC Oligochaeta; Crassiclitellata; Lumbricina; Lumbricidae; Lumbricinae;
OC Eisenia.
OX NCBI_TaxID=6396;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 45-61; 100-122; 131-149;
RP 186-205; 218-237 AND 250-277, FUNCTION, SUBCELLULAR LOCATION, AND TISSUE
RP SPECIFICITY.
RC TISSUE=Coelomocyte;
RX PubMed=9210594; DOI=10.1016/s0378-1119(97)00047-4;
RA Sekizawa Y., Kubo T., Kobayashi H., Nakajima T., Natori S.;
RT "Molecular cloning of cDNA for lysenin, a novel protein in the earthworm
RT Eisenia foetida that causes contraction of rat vascular smooth muscle.";
RL Gene 191:97-102(1997).
RN [2]
RP FUNCTION.
RX PubMed=9478988; DOI=10.1074/jbc.273.9.5300;
RA Yamaji A., Sekizawa Y., Emoto K., Sakuraba H., Inoue K., Kobayashi H.,
RA Umeda M.;
RT "Lysenin, a novel sphingomyelin-specific binding protein.";
RL J. Biol. Chem. 273:5300-5306(1998).
RN [3]
RP FUNCTION.
RX PubMed=10684578;
RX DOI=10.1002/(sici)1097-010x(20000401)286:5<538::aid-jez12>3.0.co;2-w;
RA Kobayashi H., Sekizawa Y., Aizu M., Umeda M.;
RT "Lethal and non-lethal responses of spermatozoa from a wide variety of
RT vertebrates and invertebrates to lysenin, a protein from the coelomic fluid
RT of the earthworm Eisenia foetida.";
RL J. Exp. Zool. 286:538-549(2000).
RN [4]
RP FUNCTION.
RX PubMed=12676961; DOI=10.1074/jbc.m213209200;
RA Yamaji-Hasegawa A., Makino A., Baba T., Senoh Y., Kimura-Suda H.,
RA Sato S.B., Terada N., Ohno S., Kiyokawa E., Umeda M., Kobayashi T.;
RT "Oligomerization and pore formation of a sphingomyelin-specific toxin,
RT lysenin.";
RL J. Biol. Chem. 278:22762-22770(2003).
RN [5]
RP FUNCTION, MUTAGENESIS OF TRP-20; TRP-116; TRP-187; TRP-206; TRP-245 AND
RP TRP-291, AND SITES.
RX PubMed=15274631; DOI=10.1021/bi049561j;
RA Kiyokawa E., Makino A., Ishii K., Otsuka N., Yamaji-Hasegawa A.,
RA Kobayashi T.;
RT "Recognition of sphingomyelin by lysenin and lysenin-related proteins.";
RL Biochemistry 43:9766-9773(2004).
RN [6]
RP FUNCTION.
RX PubMed=16971770; DOI=10.2220/biomedres.27.169;
RA Kobayashi H., Suzuki H., Ohta N.;
RT "Exfoliation of the epidermal cells and defecation by amphibian larvae in
RT response to coelomic fluid and lysenin from the earthworm Eisenia
RT foetida.";
RL Biomed. Res. 27:169-181(2006).
RN [7] {ECO:0000312|PDB:3ZX7, ECO:0000312|PDB:3ZXD, ECO:0000312|PDB:3ZXG}
RP X-RAY CRYSTALLOGRAPHY (2.84 ANGSTROMS) IN COMPLEX WITH SPHINGOMYELIN AND
RP ALONE, AND MUTAGENESIS OF LYS-21; TYR-24; TYR-26; GLN-117 AND GLU-128.
RX PubMed=22819216; DOI=10.1016/j.str.2012.06.011;
RA De Colibus L., Sonnen A.F., Morris K.J., Siebert C.A., Abrusci P.,
RA Plitzko J., Hodnik V., Leippe M., Volpi E., Anderluh G., Gilbert R.J.;
RT "Structures of lysenin reveal a shared evolutionary origin for pore-forming
RT proteins and its mode of sphingomyelin recognition.";
RL Structure 20:1498-1507(2012).
RN [8] {ECO:0000312|PDB:5GAQ}
RP STRUCTURE BY ELECTRON MICROSCOPY (3.14 ANGSTROMS) IN HOMO-NONAMERIC FORM,
RP AND SUBUNIT.
RX PubMed=27048994; DOI=10.1038/ncomms11293;
RA Bokori-Brown M., Martin T.G., Naylor C.E., Basak A.K., Titball R.W.,
RA Savva C.G.;
RT "Cryo-EM structure of lysenin pore elucidates membrane insertion by an
RT aerolysin family protein.";
RL Nat. Commun. 7:11293-11293(2016).
RN [9] {ECO:0000312|PDB:5EC5}
RP X-RAY CRYSTALLOGRAPHY (3.1 ANGSTROMS) IN HOMO-NONAMERIC FORM, SUBUNIT,
RP MUTAGENESIS OF VAL-88 AND TYR-131, AND BIOTECHNOLOGY.
RX PubMed=27176125; DOI=10.1038/ncomms11598;
RA Podobnik M., Savory P., Rojko N., Kisovec M., Wood N., Hambley R., Pugh J.,
RA Wallace E.J., McNeill L., Bruce M., Liko I., Allison T.M., Mehmood S.,
RA Yilmaz N., Kobayashi T., Gilbert R.J., Robinson C.V., Jayasinghe L.,
RA Anderluh G.;
RT "Crystal structure of an invertebrate cytolysin pore reveals unique
RT properties and mechanism of assembly.";
RL Nat. Commun. 7:11598-11598(2016).
CC -!- FUNCTION: Pore-forming toxin that defensively acts against parasitic
CC microorganisms by forming pores in sphingomyelin-containing membranes.
CC Has hemolytic activity and is also cytotoxic to spermatozoa of some
CC species of invertebrates and many species of vertebrates and to
CC amphibian larvae, guinea pig polymorphonuclear leukocytes, chicken
CC fibroblasts, normal spleen cells and various tumor cells. Is lethal for
CC various species of reptiles, amphibian, birds and mammals. Induces
CC smooth muscle contraction. It binds sphingomyelin and induces hemolysis
CC in the same manner as lysenin-related protein 2, and is 10-fold more
CC effective than lysenin-related protein 1. {ECO:0000269|PubMed:10684578,
CC ECO:0000269|PubMed:12676961, ECO:0000269|PubMed:15274631,
CC ECO:0000269|PubMed:16971770, ECO:0000269|PubMed:9210594,
CC ECO:0000269|PubMed:9478988}.
CC -!- SUBUNIT: Binds to sphingomyelin as a monomer by using its C-terminal
CC domain (PubMed:22819216). Forms a nonamer when sphingomyelin/lysenin
CC ratio is lower than ca 500 (PubMed:27048994, PubMed:27176125).
CC Oligomerization, but not binding, is influenced by the fluidity of
CC sphingomyelin. {ECO:0000269|PubMed:22819216,
CC ECO:0000269|PubMed:27176125}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:9210594}. Target
CC cell membrane {ECO:0000269|PubMed:9210594}. Note=Forms a beta-barrel
CC pore in the membrane. {ECO:0000269|PubMed:27048994,
CC ECO:0000269|PubMed:27176125}.
CC -!- TISSUE SPECIFICITY: Expressed by coelomocytes.
CC {ECO:0000269|PubMed:9210594}.
CC -!- BIOTECHNOLOGY: This toxin represents an excellent tool for visualizing
CC distribution and dynamics of sphingomyelin in cells.
CC {ECO:0000305|PubMed:27176125}.
CC -!- SIMILARITY: Belongs to the lysenin family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; D85846; BAA21518.1; -; mRNA.
DR PDB; 3ZX7; X-ray; 2.84 A; A=2-297.
DR PDB; 3ZXD; X-ray; 3.30 A; A/B/C/D=2-297.
DR PDB; 3ZXG; X-ray; 3.12 A; A/B=2-297.
DR PDB; 5EC5; X-ray; 3.10 A; A/B/C/D/E/F/G/H/I/J/K/L/M/N/O/P/R/S=1-297.
DR PDB; 5GAQ; EM; 3.14 A; A/B/C/D/E/F/G/H/I=1-297.
DR PDBsum; 3ZX7; -.
DR PDBsum; 3ZXD; -.
DR PDBsum; 3ZXG; -.
DR PDBsum; 5EC5; -.
DR PDBsum; 5GAQ; -.
DR AlphaFoldDB; O18423; -.
DR SMR; O18423; -.
DR DIP; DIP-59857N; -.
DR TCDB; 1.C.43.1.1; the earthworm lysenin toxin (lysenin) family.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0042742; P:defense response to bacterium; IEA:UniProtKB-KW.
DR GO; GO:0044179; P:hemolysis in another organism; IEA:UniProtKB-KW.
DR GO; GO:0006811; P:ion transport; IEA:UniProtKB-KW.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic; Antimicrobial; Cytolysis;
KW Direct protein sequencing; Disulfide bond; Hemolysis; Ion transport;
KW Membrane; Secreted; Target cell membrane; Target membrane; Toxin;
KW Transmembrane; Transport.
FT CHAIN 1..297
FT /note="Lysenin"
FT /evidence="ECO:0000305|PubMed:9210594"
FT /id="PRO_0000342606"
FT REGION 10..33
FT /note="N-terminal cap domain"
FT /evidence="ECO:0000305|PubMed:27048994"
FT REGION 34..107
FT /note="Beta-hairpin domain"
FT /evidence="ECO:0000305|PubMed:27048994"
FT REGION 108..156
FT /note="N-terminal cap domain"
FT /evidence="ECO:0000305|PubMed:27048994"
FT REGION 157..297
FT /note="C-terminal receptor-binding domain"
FT /evidence="ECO:0000305|PubMed:27048994"
FT BINDING 185
FT /ligand="an N-(acyl)-sphingosylphosphocholine"
FT /ligand_id="ChEBI:CHEBI:64583"
FT /evidence="ECO:0000269|PubMed:22819216"
FT BINDING 227
FT /ligand="an N-(acyl)-sphingosylphosphocholine"
FT /ligand_id="ChEBI:CHEBI:64583"
FT /evidence="ECO:0000269|PubMed:22819216"
FT BINDING 233
FT /ligand="an N-(acyl)-sphingosylphosphocholine"
FT /ligand_id="ChEBI:CHEBI:64583"
FT /evidence="ECO:0000269|PubMed:22819216"
FT BINDING 282
FT /ligand="an N-(acyl)-sphingosylphosphocholine"
FT /ligand_id="ChEBI:CHEBI:64583"
FT /evidence="ECO:0000269|PubMed:22819216"
FT SITE 20
FT /note="Crucial for binding sphingomyelin and inducing
FT hemolysis"
FT /evidence="ECO:0000269|PubMed:15274631"
FT SITE 187
FT /note="Crucial for binding sphingomyelin and important for
FT inducing hemolysis"
FT /evidence="ECO:0000269|PubMed:15274631"
FT SITE 209
FT /note="Important for activity"
FT SITE 245
FT /note="Crucial for binding sphingomyelin and inducing
FT hemolysis"
FT /evidence="ECO:0000269|PubMed:15274631"
FT SITE 291
FT /note="Crucial for binding sphingomyelin and inducing
FT hemolysis"
FT /evidence="ECO:0000269|PubMed:15274631"
FT DISULFID 272..283
FT /evidence="ECO:0000255"
FT MUTAGEN 20
FT /note="W->A: Loss of ability to bind sphingomyelin, and to
FT induce hemolysis."
FT /evidence="ECO:0000269|PubMed:15274631"
FT MUTAGEN 21
FT /note="K->A: Decrease in ability to bind sphingomyelin and
FT to lyse cells."
FT /evidence="ECO:0000269|PubMed:22819216"
FT MUTAGEN 24
FT /note="Y->A: In double Tyr mutant; almost complete loss of
FT ability to bind sphingomyelin and to lyse cells."
FT /evidence="ECO:0000269|PubMed:22819216"
FT MUTAGEN 26
FT /note="Y->A: In double Tyr mutant; almost complete loss of
FT ability to bind sphingomyelin and to lyse cells."
FT /evidence="ECO:0000269|PubMed:22819216"
FT MUTAGEN 88
FT /note="V->C: In double Cys mutant; complete loss of ability
FT to form pores when Cys are disulfide-linked."
FT /evidence="ECO:0000269|PubMed:27176125"
FT MUTAGEN 117
FT /note="Q->A: Decrease in ability to bind sphingomyelin and
FT to lyse cells."
FT /evidence="ECO:0000269|PubMed:22819216"
FT MUTAGEN 128
FT /note="E->A: Decrease in ability to bind sphingomyelin and
FT to lyse cells."
FT /evidence="ECO:0000269|PubMed:22819216"
FT MUTAGEN 131
FT /note="Y->C: In double Cys mutant; complete loss of ability
FT to form pores when Cys are disulfide-linked."
FT /evidence="ECO:0000269|PubMed:27176125"
FT MUTAGEN 187
FT /note="W->A: Loss of ability to bind sphingomyelin, and
FT induces hemolysis only at high concentration."
FT /evidence="ECO:0000269|PubMed:15274631"
FT MUTAGEN 206
FT /note="W->A: Does not affect binding, and has little loss
FT of hemolytic activity."
FT /evidence="ECO:0000269|PubMed:15274631"
FT MUTAGEN 245
FT /note="W->A: Loss of ability to bind sphingomyelin, and to
FT induce hemolysis."
FT /evidence="ECO:0000269|PubMed:15274631"
FT MUTAGEN 291
FT /note="W->A: Loss of ability to bind sphingomyelin, and to
FT induce hemolysis."
FT /evidence="ECO:0000269|PubMed:15274631"
FT STRAND 9..15
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 17..28
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 30..32
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 34..44
FT /evidence="ECO:0007829|PDB:3ZX7"
FT TURN 61..63
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 69..80
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 82..90
FT /evidence="ECO:0007829|PDB:3ZX7"
FT HELIX 92..95
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 99..107
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 111..122
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 125..132
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 136..138
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 147..158
FT /evidence="ECO:0007829|PDB:3ZX7"
FT TURN 163..165
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 167..172
FT /evidence="ECO:0007829|PDB:3ZX7"
FT TURN 173..176
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 177..184
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 187..194
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 200..205
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 208..214
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 217..220
FT /evidence="ECO:0007829|PDB:5GAQ"
FT STRAND 223..226
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 231..235
FT /evidence="ECO:0007829|PDB:3ZX7"
FT HELIX 241..243
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 245..249
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 257..259
FT /evidence="ECO:0007829|PDB:5GAQ"
FT STRAND 261..266
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 269..273
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 281..285
FT /evidence="ECO:0007829|PDB:3ZX7"
FT STRAND 291..295
FT /evidence="ECO:0007829|PDB:3ZX7"
SQ SEQUENCE 297 AA; 33441 MW; B9D06B3543CD48F0 CRC64;
MSAKAAEGYE QIEVDVVAVW KEGYVYENRG STSVDQKITI TKGMKNVNSE TRTVTATHSI
GSTISTGDAF EIGSVEVSYS HSHEESQVSM TETEVYESKV IEHTITIPPT SKFTRWQLNA
DVGGADIEYM YLIDEVTPIG GTQSIPQVIT SRAKIIVGRQ IILGKTEIRI KHAERKEYMT
VVSRKSWPAA TLGHSKLFKF VLYEDWGGFR IKTLNTMYSG YEYAYSSDQG GIYFDQGTDN
PKQRWAINKS LPLRHGDVVT FMNKYFTRSG LCYDDGPATN VYCLDKREDK WILEVVG
