PA2H2_BOTAS
ID PA2H2_BOTAS Reviewed; 137 AA.
AC P24605;
DT 01-MAR-1992, integrated into UniProtKB/Swiss-Prot.
DT 15-JAN-2008, sequence version 3.
DT 03-AUG-2022, entry version 113.
DE RecName: Full=Basic phospholipase A2 homolog 2;
DE Short=svPLA2 homolog;
DE AltName: Full=Myotoxin II {ECO:0000303|PubMed:2781572, ECO:0000303|PubMed:9654096};
DE Short=Basp-II {ECO:0000303|PubMed:15961104};
DE Short=Lys49 Mt-II {ECO:0000303|PubMed:20660736};
DE Short=MtxII;
DE Flags: Precursor;
OS Bothrops asper (Terciopelo).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
OC Lepidosauria; Squamata; Bifurcata; Unidentata; Episquamata; Toxicofera;
OC Serpentes; Colubroidea; Viperidae; Crotalinae; Bothrops.
OX NCBI_TaxID=8722;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA].
RC TISSUE=Venom gland;
RX PubMed=11240369; DOI=10.1016/s1357-2725(00)00073-x;
RA Lizano S., Lambeau G., Lazdunski M.;
RT "Cloning and cDNA sequence analysis of Lys(49) and Asp(49) basic
RT phospholipase A(2) myotoxin isoforms from Bothrops asper.";
RL Int. J. Biochem. Cell Biol. 33:127-132(2001).
RN [2]
RP PROTEIN SEQUENCE OF 17-137, FUNCTION, SUBUNIT, SUBCELLULAR LOCATION, AND
RP VARIANT PHE-130.
RC STRAIN=Caribbean; TISSUE=Venom;
RX PubMed=1899180; DOI=10.1016/0003-9861(91)90307-5;
RA Francis B., Gutierrez J.M., Lomonte B., Kaiser I.I.;
RT "Myotoxin II from Bothrops asper (Terciopelo) venom is a lysine-49
RT phospholipase A2.";
RL Arch. Biochem. Biophys. 284:352-359(1991).
RN [3]
RP PROTEIN SEQUENCE OF 125-132, MASS SPECTROMETRY, SUBCELLULAR LOCATION, AND
RP VARIANT PHE-130.
RC STRAIN=Caribbean, and Pacific; TISSUE=Venom;
RX PubMed=35248586; DOI=10.1016/j.toxicon.2022.02.024;
RA Lomonte B., Fernandez J.;
RT "Solving the microheterogeneity of Bothrops asper myotoxin-II by high-
RT resolution mass spectrometry: insights into C-terminal region variability
RT in Lys49-phospholipase A2 homologs.";
RL Toxicon 210:123-131(2022).
RN [4]
RP AMINO-ACID COMPOSITION, FUNCTION, AND SUBUNIT.
RC TISSUE=Venom;
RX PubMed=2781572; DOI=10.1016/0041-0101(89)90039-1;
RA Lomonte B., Gutierrez J.M.;
RT "A new muscle damaging toxin, myotoxin II, from the venom of the snake
RT Bothrops asper (terciopelo).";
RL Toxicon 27:725-733(1989).
RN [5]
RP FUNCTION, ACTIVITY REGULATION, SYNTHESIS OF 121-133, AND SUBUNIT.
RC TISSUE=Venom;
RX PubMed=7961981; DOI=10.1016/s0021-9258(18)43961-0;
RA Lomonte B., Moreno E., Tarkowski A., Hanson L.A., Maccarana M.;
RT "Neutralizing interaction between heparins and myotoxin II, a lysine 49
RT phospholipase A2 from Bothrops asper snake venom. Identification of a
RT heparin-binding and cytolytic toxin region by the use of synthetic peptides
RT and molecular modeling.";
RL J. Biol. Chem. 269:29867-29873(1994).
RN [6]
RP FUNCTION ON A VARIETY OF CELL TYPES.
RX PubMed=7886694; DOI=10.1016/0041-0101(94)90408-1;
RA Lomonte B., Tarkowski A., Hanson L.A.;
RT "Broad cytolytic specificity of myotoxin II, a lysine-49 phospholipase A2
RT of Bothrops asper snake venom.";
RL Toxicon 32:1359-1369(1994).
RN [7]
RP SUBUNIT.
RX PubMed=7718570; DOI=10.1021/bi00014a021;
RA Pedersen J.Z., Lomonte B., Massoud R., Gubensek F., Gutierrez J.M.,
RA Rufini S.;
RT "Autocatalytic acylation of phospholipase-like myotoxins.";
RL Biochemistry 34:4670-4675(1995).
RN [8]
RP FUNCTION ON BACTERIA, AND SYNTHESIS OF 121-133.
RC TISSUE=Venom;
RX PubMed=9654096; DOI=10.1046/j.1432-1327.1998.2530452.x;
RA Paramo L., Lomonte B., Pizarro-Cerda J., Bengoechea J.A., Gorvel J.P.,
RA Moreno E.;
RT "Bactericidal activity of Lys49 and Asp49 myotoxic phospholipases A2 from
RT Bothrops asper snake venom--synthetic Lys49 myotoxin II-(115-129)-peptide
RT identifies its bactericidal region.";
RL Eur. J. Biochem. 253:452-461(1998).
RN [9]
RP FUNCTION IN EDEMA.
RX PubMed=9839670; DOI=10.1016/s0041-0101(98)00107-x;
RA Chaves F., Leon G., Alvarado V.H., Gutierrez J.M.;
RT "Pharmacological modulation of edema induced by Lys-49 and Asp-49 myotoxic
RT phospholipases A2 isolated from the venom of the snake Bothrops asper
RT (terciopelo).";
RL Toxicon 36:1861-1869(1998).
RN [10]
RP FUNCTION IN CYTOTOXICITY.
RC TISSUE=Venom;
RX PubMed=9920486; DOI=10.1016/s0041-0101(98)00171-8;
RA Lomonte B., Angulo Y., Rufini S., Cho W., Giglio J.R., Ohno M.,
RA Daniele J.J., Geoghegan P., Gutierrez J.M.;
RT "Comparative study of the cytolytic activity of myotoxic phospholipases A2
RT on mouse endothelial (tEnd) and skeletal muscle (C2C12) cells in vitro.";
RL Toxicon 37:145-158(1999).
RN [11]
RP FUNCTION AS AN ANTICOAGULANT.
RX PubMed=18062812; DOI=10.1186/1472-6807-7-82;
RA Faure G., Gowda V.T., Maroun R.C.;
RT "Characterization of a human coagulation factor Xa-binding site on
RT Viperidae snake venom phospholipases A2 by affinity binding studies and
RT molecular bioinformatics.";
RL BMC Struct. Biol. 7:82-82(2007).
RN [12]
RP FUNCTION.
RC TISSUE=Venom;
RX PubMed=20660736; DOI=10.1073/pnas.1009128107;
RA Cintra-Francischinelli M., Caccin P., Chiavegato A., Pizzo P.,
RA Carmignoto G., Angulo Y., Lomonte B., Gutierrez J.M., Montecucco C.;
RT "Bothrops snake myotoxins induce a large efflux of ATP and potassium with
RT spreading of cell damage and pain.";
RL Proc. Natl. Acad. Sci. U.S.A. 107:14140-14145(2010).
RN [13]
RP ACTIVITY REGULATION BY MEMBRANE CHOLESTEROL, BIOPHYSICOCHEMICAL PROPERTIES,
RP AND SYNTHESIS OF 121-133.
RC TISSUE=Venom;
RX PubMed=21506137; DOI=10.1002/cbf.1758;
RA Rangel J., Quesada O., Gutierrez J.M., Angulo Y., Lomonte B.;
RT "Membrane cholesterol modulates the cytolytic mechanism of myotoxin II, a
RT Lys49 phospholipase A2 homologue from the venom of Bothrops asper.";
RL Cell Biochem. Funct. 29:365-370(2011).
RN [14]
RP PHARMACEUTICAL, AND SYNTHESIS OF 17-36.
RX PubMed=26931059; DOI=10.1038/srep22413;
RA Smith A.I., Rajapakse N.W., Kleifeld O., Lomonte B., Sikanyika N.L.,
RA Spicer A.J., Hodgson W.C., Conroy P.J., Small D.H., Kaye D.M.,
RA Parkington H.C., Whisstock J.C., Kuruppu S.;
RT "N-terminal domain of Bothrops asper Myotoxin II enhances the activity of
RT endothelin converting enzyme-1 and neprilysin.";
RL Sci. Rep. 6:22413-22413(2016).
RN [15]
RP ERRATUM OF PUBMED:26931059.
RX PubMed=27102936; DOI=10.1038/srep24333;
RA Smith A.I., Rajapakse N.W., Kleifeld O., Lomonte B., Sikanyika N.L.,
RA Spicer A.J., Hodgson W.C., Conroy P.J., Small D.H., Kaye D.M.,
RA Parkington H.C., Whisstock J.C., Kuruppu S.;
RT "Corrigendum: N-terminal domain of Bothrops asper Myotoxin II enhances the
RT activity of endothelin converting enzyme-1 and neprilysin.";
RL Sci. Rep. 6:24333-24333(2016).
RN [16] {ECO:0000312|PDB:1CLP}
RP X-RAY CRYSTALLOGRAPHY (2.80 ANGSTROMS) OF 17-137, DISULFIDE BONDS, AND
RP SUBUNIT.
RC TISSUE=Venom;
RX PubMed=15299297; DOI=10.1107/s0907444994011455;
RA Arni R.K., Ward R.J., Gutierrez J.M., Tulinsky A.;
RT "Structure of a calcium-independent phospholipase-like myotoxic protein
RT from Bothrops asper venom.";
RL Acta Crystallogr. D 51:311-317(1995).
RN [17] {ECO:0000312|PDB:1Y4L}
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 17-137 IN COMPLEX WITH SURAMIN,
RP ACTIVITY REGULATION, AND DISULFIDE BONDS.
RC TISSUE=Venom;
RX PubMed=15961104; DOI=10.1016/j.jmb.2005.04.072;
RA Murakami M.T., Arruda E.Z., Melo P.A., Martinez A.B., Calil-Elias S.,
RA Tomaz M.A., Lomonte B., Gutierrez J.M., Arni R.K.;
RT "Inhibition of myotoxic activity of Bothrops asper myotoxin II by the anti-
RT trypanosomal drug suramin.";
RL J. Mol. Biol. 350:416-426(2005).
CC -!- FUNCTION: Snake venom phospholipase A2 homolog that lacks enzymatic
CC activity. Is myotoxic and induces a dose-dependent edema in the mouse
CC foot pad (PubMed:2781572, PubMed:9839670). Also exhibits strong
CC anticoagulant effects by binding to factor Xa (F10) and inhibiting the
CC prothrombinase activity (IC(50) is 3 nM) (PubMed:18062812). In
CC addition, it shows cytotoxic activity to a variety of cell types and
CC bactericidal activity to a variety of Gram-negative and Gram-positive
CC bacteria (PubMed:7886694, PubMed:9654096, PubMed:9920486). Also induces
CC a very rapid release of large amounts of potassium ions and ATP from
CC muscle cells, which accounts for the pain reaction characteristic of
CC viperid envenomations (PubMed:20660736). The released ATP amplifies the
CC effect of the myotoxins, acting as a 'danger signal', which spreads and
CC causes further damage by acting on purinergic receptors
CC (PubMed:20660736). A model of myotoxic mechanism has been proposed: an
CC apo Lys49-PLA2 is activated by the entrance of a hydrophobic molecule
CC (e.g. fatty acid) at the hydrophobic channel of the protein leading to
CC a reorientation of a monomer (By similarity). This reorientation causes
CC a transition between 'inactive' to 'active' states, causing alignment
CC of C-terminal and membrane-docking sites (MDoS) side-by-side and
CC putting the membrane-disruption sites (MDiS) in the same plane, exposed
CC to solvent and in a symmetric position for both monomers (By
CC similarity). The MDoS region stabilizes the toxin on membrane by the
CC interaction of charged residues with phospholipid head groups (By
CC similarity). Subsequently, the MDiS region destabilizes the membrane
CC with penetration of hydrophobic residues (By similarity). This
CC insertion causes a disorganization of the membrane, allowing an
CC uncontrolled influx of ions (i.e. calcium and sodium), and eventually
CC triggering irreversible intracellular alterations and cell death (By
CC similarity). {ECO:0000250|UniProtKB:I6L8L6,
CC ECO:0000269|PubMed:18062812, ECO:0000269|PubMed:1899180,
CC ECO:0000269|PubMed:20660736, ECO:0000269|PubMed:2781572,
CC ECO:0000269|PubMed:7886694, ECO:0000269|PubMed:9654096,
CC ECO:0000269|PubMed:9839670, ECO:0000269|PubMed:9920486}.
CC -!- ACTIVITY REGULATION: Heparin inhibits the myotoxic activity
CC (PubMed:7961981). Suramin inhibits the myotoxic activity
CC (PubMed:15961104). High level of membrane cholesterol content reduces
CC cytolytic activity, whereas low level of membrane cholesterol content
CC increases cytolytic activity (PubMed:21506137).
CC {ECO:0000269|PubMed:15961104, ECO:0000269|PubMed:21506137,
CC ECO:0000269|PubMed:7961981}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Temperature dependence:
CC Cytolytic activity upon C2C12 cells is completely abolished at 15
CC degrees Celsius or below. {ECO:0000269|PubMed:21506137};
CC -!- SUBUNIT: Homodimer; non-covalently linked (PubMed:1899180,
CC PubMed:2781572, PubMed:7718570, PubMed:15299297). Binds to heparin
CC (PubMed:7961981). {ECO:0000269|PubMed:15299297,
CC ECO:0000269|PubMed:1899180, ECO:0000269|PubMed:2781572,
CC ECO:0000269|PubMed:7718570}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:1899180}.
CC -!- TISSUE SPECIFICITY: Expressed by the venom gland.
CC {ECO:0000305|PubMed:1899180}.
CC -!- PTM: It binds long-chain fatty acids covalently by a rapid,
CC spontaneous, and autocatalytic process. When acylated, it binds to the
CC surface of liposomes and isolated muscle membranes, with the fatty acid
CC moiety inserted into the lipid bilayer and possibly acting as an anchor
CC (PubMed:7718570). {ECO:0000269|PubMed:7718570}.
CC -!- MASS SPECTROMETRY: Mass=13749.6; Method=Electrospray; Note=snakes from
CC Pacific region, Monoisotopic mass.;
CC Evidence={ECO:0000269|PubMed:35248586};
CC -!- MASS SPECTROMETRY: Mass=13716.5; Method=Electrospray; Note=snakes from
CC Caribbean region, Monoisotopic mass.;
CC Evidence={ECO:0000269|PubMed:35248586};
CC -!- PHARMACEUTICAL: K49-P1-20, a twenty-residue N-terminal peptide that
CC corresponds to residues 17-36 of myotoxin II, enhances the activity of
CC endothelin converting enzyme-1 (ECE1) and neprilysin (MME), two enzymes
CC that degrade amyloid beta in the brain. It could be an excellent
CC research tool to study mechanisms of ECE1 and MME stimulation, and a
CC potential novel drug lead in the fight against Alzheimer's disease.
CC {ECO:0000305|PubMed:26931059}.
CC -!- SIMILARITY: Belongs to the phospholipase A2 family. Group II subfamily.
CC K49 sub-subfamily. {ECO:0000305}.
CC -!- CAUTION: Does not bind calcium as one of the calcium-binding sites is
CC lost (Asp->Lys in position 64, which corresponds to 'Lys-49' in the
CC current nomenclature). {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 PIR; S13900; S13900.
DR PDB; 1CLP; X-ray; 2.80 A; A/B=17-137.
DR PDB; 1Y4L; X-ray; 1.70 A; A/B=17-137.
DR PDBsum; 1CLP; -.
DR PDBsum; 1Y4L; -.
DR AlphaFoldDB; P24605; -.
DR SMR; P24605; -.
DR Allergome; 6319; Bot as 1.
DR EvolutionaryTrace; P24605; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0005509; F:calcium ion binding; IEA:InterPro.
DR GO; GO:0008201; F:heparin binding; IEA:UniProtKB-KW.
DR GO; GO:0004623; F:phospholipase A2 activity; IEA:InterPro.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0050482; P:arachidonic acid secretion; IEA:InterPro.
DR GO; GO:0042742; P:defense response to bacterium; IEA:UniProtKB-KW.
DR GO; GO:0016042; P:lipid catabolic process; IEA:InterPro.
DR GO; GO:0006644; P:phospholipid metabolic process; IEA:InterPro.
DR CDD; cd00125; PLA2c; 1.
DR Gene3D; 1.20.90.10; -; 1.
DR InterPro; IPR001211; PLipase_A2.
DR InterPro; IPR033112; PLipase_A2_Asp_AS.
DR InterPro; IPR016090; PLipase_A2_dom.
DR InterPro; IPR036444; PLipase_A2_dom_sf.
DR InterPro; IPR033113; PLipase_A2_His_AS.
DR PANTHER; PTHR11716; PTHR11716; 1.
DR Pfam; PF00068; Phospholip_A2_1; 1.
DR PRINTS; PR00389; PHPHLIPASEA2.
DR SMART; SM00085; PA2c; 1.
DR SUPFAM; SSF48619; SSF48619; 1.
DR PROSITE; PS00119; PA2_ASP; 1.
DR PROSITE; PS00118; PA2_HIS; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic; Antimicrobial;
KW Blood coagulation cascade inhibiting toxin; Direct protein sequencing;
KW Disulfide bond; Hemostasis impairing toxin; Heparin-binding; Lipoprotein;
KW Myotoxin; Pharmaceutical; Secreted; Signal; Toxin.
FT SIGNAL 1..16
FT /evidence="ECO:0000269|PubMed:1899180"
FT CHAIN 17..137
FT /note="Basic phospholipase A2 homolog 2"
FT /id="PRO_0000161618"
FT REGION 121..133
FT /note="Important for membrane-damaging activities in
FT eukaryotes and bacteria; heparin-binding"
FT /evidence="ECO:0000269|PubMed:21506137,
FT ECO:0000269|PubMed:7961981, ECO:0000269|PubMed:9654096"
FT SITE 121
FT /note="Important residue of the cationic membrane-docking
FT site (MDoS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT SITE 124
FT /note="Important residue of the cationic membrane-docking
FT site (MDoS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT SITE 127
FT /note="Hydrophobic membrane-disruption site (MDiS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT SITE 128
FT /note="Cationic membrane-docking site (MDoS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT SITE 130
FT /note="Hydrophobic membrane-disruption site (MDiS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT SITE 133
FT /note="Cationic membrane-docking site (MDoS)"
FT /evidence="ECO:0000250|UniProtKB:I6L8L6"
FT DISULFID 42..131
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 44..60
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 59..111
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 65..137
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 66..104
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 73..97
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT DISULFID 91..102
FT /evidence="ECO:0000269|PubMed:15299297,
FT ECO:0000269|PubMed:15961104, ECO:0000312|PDB:1CLP,
FT ECO:0000312|PDB:1Y4L"
FT VARIANT 130
FT /note="L -> F (leu in snakes from Caribbean region, Phe in
FT snakes from Pacific region)"
FT /evidence="ECO:0000269|PubMed:1899180,
FT ECO:0000269|PubMed:35248586"
FT HELIX 18..29
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 33..37
FT /evidence="ECO:0007829|PDB:1Y4L"
FT TURN 41..43
FT /evidence="ECO:0007829|PDB:1Y4L"
FT STRAND 44..47
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 55..67
FT /evidence="ECO:0007829|PDB:1Y4L"
FT TURN 75..77
FT /evidence="ECO:0007829|PDB:1Y4L"
FT STRAND 82..85
FT /evidence="ECO:0007829|PDB:1Y4L"
FT STRAND 88..91
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 96..114
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 116..118
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 121..123
FT /evidence="ECO:0007829|PDB:1Y4L"
FT HELIX 128..130
FT /evidence="ECO:0007829|PDB:1Y4L"
SQ SEQUENCE 137 AA; 15509 MW; F7194B8E794D0C3D CRC64;
MRTLWIMAVL LVGVEGSLFE LGKMILQETG KNPAKSYGAY GCNCGVLGRG KPKDATDRCC
YVHKCCYKKL TGCNPKKDRY SYSWKDKTIV CGENNSCLKE LCECDKAVAI CLRENLNTYN
KKYRYYLKPL CKKADAC
