Supplementary Data File 1: Clustering and phylogenetic analysis of Acropora digitifera toxins
HMM based hierarchical clustering (HHCompare):
HHCompare clustering was performed at HMM-HMM similarity e-value of 1.0e-20. Following the
clustering, sequences within each group were aligned using MUSCLE. For three sequence groups,
phylogentic trees were constructed using Minimal Evolution method, while larger groups were
analyzed using Maximum Likelihood (ML) method. In case of ML, evolutionary model was inferred by
MEGA 6.0 model selection tool, based on Sample-corrected Akaike information criterion (AICc).
adi v1.07444 F Astacin-like metalloprotease toxin 3 OS=Loxosceles intermedia
adi v1.15751 F Astacin-like metalloprotease toxin 1 OS=Loxosceles intermedia
adi v1.17845 F Nematocyte expressed protein 6 OS=Nematostella vectensis
Group 1
adi v1.20368 T Nematocyte expressed protein 6 OS=Nematostella vectensis
adi v1.13648 F Nematocyte expressed protein 6 OS=Nematostella vectensis
adi v1.23604 T Astacin-like metalloprotease toxin 2 OS=Loxosceles intermedia
adi v1.15074 F Venom dipeptidyl peptidase 4 OS=Vespula vulgaris
Group 2
adi v1.20292 T Venom dipeptidyl peptidase 4 OS=Vespula vulgaris
adi v1.02125 T Venom phosphodiesterase 2 OS=Crotalus adamanteus
Group 3
adi v1.16452 F Venom phosphodiesterase 2 OS=Crotalus adamanteus
adi v1.08969 T Phospholipase-B 81 OS=Drysdalia coronoides
Group 4
adi v1.12172 T Phospholipase-B 81 OS=Drysdalia coronoides
adi v1.11797 F Neutral phospholipase A2 homolog taipoxin beta chain 1 OS=Oxyuranus scutellatus scutellatus
adi v1.16921 F Phospholipase A2 OS=Condylactis gigantea
adi v1.09427 F delta-TATX-Avl2a
adi v1.14353 T delta-ALTX-Pse2b
Group 6
adi v1.16619 F delta-TATX-Avl2a
adi v1.11218 F Kunitz-type serine protease inhibitor mulgin-3 OS=Pseudechis australis
Group 7
adi v1.23374 T Kunitz-type serine protease inhibitor Hg1 OS=Hadrurus gertschi
adi v1.16440 F delta-AITX-Aas1a
Group 8
adi v1.24162 F Equinatoxin-2 OS=Actinia equina
adi v1.01810 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS=Crotalus adamanteus
adi v1.04835 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS=Crotalus adamanteus
Group 9
adi v1.23821 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS=Crotalus adamanteus
adi v1.00020 F Putative endothelial lipase OS=Crotalus adamanteus
adi v1.09322 F Putative endothelial lipase OS=Crotalus adamanteus
Group 10
adi v1.12434 F Phospholipase A1 OS=Polybia paulista
adi v1.08904 T Blarina toxin OS=Blarina brevicauda GN=BTX
adi v1.16374 T Venom prothrombin activator notecarin-D2 OS=Notechis scutatus scutatus
adi v1.15821 F Coagulation factor X OS=Tropidechis carinatus GN
Group 11
adi v1.09733 T Venom peptide isomerase heavy chain OS=Agelenopsis aperta
adi v1.23657 F Venom peptide isomerase heavy chain OS=Agelenopsis aperta
adi v1.01092 T Translationally-controlled tumor protein homolog OS=Crotalus adamanteus
adi v1.01102 T Calglandulin OS=Tropidechis carinatus
adi v1.03437 F Calglandulin OS=Tropidechis carinatus
adi v1.05162 T Reticulocalbin-2 OS=Crotalus adamanteus
adi v1.05180 T Venom carboxylesterase-6 OS=Apis mellifera
adi v1.05505 F Venom serine carboxypeptidase OS=Apis mellifera
adi v1.06821 F Hyaluronidase-1 OS=Bitis arietans
adi v1.06850 F Millepora cytotoxin-1 OS=Millepora dichotoma
adi v1.09601 T Equinatoxin-5 OS=Actinia equina
adi v1.09855 T SE-cephalotoxin OS=Sepia esculenta
adi v1.10410 F delta-AITX-Ucs1a
adi v1.10508 T Putative protein-glutamate O-methyltransferase OS=Pimpla hypochondriaca GN=vpr2
adi v1.12125 F Venom factor OS=Crotalus adamanteus
adi v1.12298 T Snaclec 7 OS=Daboia siamensis
adi v1.12311 F Toxin CfTX-1 OS=Chironex fleckeri
adi v1.14946 F Snake venom 5-nucleotidase OS=Gloydius brevicaudus
adi v1.16469 F L-amino-acid oxidase OS=Ophiophagus hannah
adi v1.18628 F Conodipine-M alpha chain OS=Conus magus
adi v1.18989 F Natterin-4 OS=Thalassophryne nattereri
adi v1.19322 F Venom allergen 3 OS=Solenopsis richteri
adi v1.19445 F Snake venom metalloprotease OS=Philodryas olfersii
adi v1.19802 F C-type lectin lectoxin-Thr1 OS=Thrasops jacksonii
adi v1.22096 F Ryncolin-4 OS=Cerberus rynchops
Figure 1. HMM-based hierarchical clustering of coral toxins. Each split indicates HMM-HMM
similarity with e-value below 1.0e-20.
Following the HMM-based clustering, phylogenetic trees were constructed for groups with 3 or
more sequences.
Group 5
adi v1.07444 F Astacin-like metalloprotease toxin 3 OS Loxosceles intermedia
adi v1.15751 F Astacin-like metalloprotease toxin 1 OS Loxosceles intermedia
adi v1.17845 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.20368 T Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.13648 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.23604 T Astacin-like metalloprotease toxin 2 OS Loxosceles intermedia
A: Group 1, ML analysis using LG+G model with 4 discrete gamma categories
adi v1.09427 F delta-TATX-Avl2a
adi v1.14353 T delta-ALTX-Pse2b
adi v1.16619 F delta-TATX-Avl2a
B: Group 6, Minimal Evolution analysis
0.2
adi v1.01810 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.04835 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.23821 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
0.1
C: Group 9, Minimal Evolution analysis
adi v1.00020 F Putative endothelial lipase OS Crotalus adamanteus
adi v1.09322 F Putative endothelial lipase OS Crotalus adamanteus
adi v1.12434 F Phospholipase A1 OS Polybia paulista
0.1
D: Group 10, Minimal Evolution analysis
adi v1.08904 T Blarina toxin OS Blarina brevicauda GN BTX
adi v1.16374 T Venom prothrombin activator notecarin-D2 OS Notechis scutatus scutatus
adi v1.23657 F Venom peptide isomerase heavy chain OS Agelenopsis aperta
adi v1.15821 F Coagulation factor X OS Tropidechis carinatus GN
adi v1.09733 T Venom peptide isomerase heavy chain OS Agelenopsis aperta
0.2
E: Group 11, ML analysis using JTT model with 4 discrete gamma categories
Figure 2. Phylogenetic analysis of HMM clustered groups
Maximium likelihood (ML) clustering:
ML based clustering was performed using MEGA 6.0. Sequences were aligned using MUSCLE,
JTT+Gamma (4 discrete gamma categories) evolutionary model was determined by MEGA 6.0 model
selection tool based on AICc, and reconstruction performed by ML method, using all amino-acids in
the alignment and 100 bootstraps. Tree was condensed based on bootstrap cutoff of 35.
adi v1.00020 F Putative endothelial lipase OS Crotalus adamanteus
Group 10
adi v1.09322 F Putative endothelial lipase OS Crotalus adamanteus
adi v1.12434 F Phospholipase A1 OS Polybia paulista
adi v1.08904 T Blarina toxin OS Blarina brevicauda GN BTX
Group 11-1
adi v1.15821 F Coagulation factor X OS Tropidechis carinatus GN
adi v1.16374 T Venom prothrombin activator notecarin-D2 OS Notechis scutatus scutatus
adi v1.19802 F C-type lectin lectoxin-Thr1 OS Thrasops jacksonii
adi v1.09733 T Venom peptide isomerase heavy chain OS Agelenopsis aperta
Group 11-2
adi v1.23657 F Venom peptide isomerase heavy chain OS Agelenopsis aperta
adi v1.01092 T Translationally-controlled tumor protein homolog OS Crotalus adamanteus
adi v1.16452 F Venom phosphodiesterase 2 OS Crotalus adamanteus
adi v1.02125 T Venom phosphodiesterase 2 OS Crotalus adamanteus
adi v1.12298 T Snaclec 7 OS Daboia siamensis
adi v1.06821 F Hyaluronidase-1 OS Bitis arietans
adi v1.08969 T Phospholipase-B 81 OS Drysdalia coronoides
Group 4
adi v1.12172 T Phospholipase-B 81 OS Drysdalia coronoides
adi v1.23604 T Astacin-like metalloprotease toxin 2 OS Loxosceles intermedia
adi v1.09601 T Equinatoxin-5 OS Actinia equina
Group 1
Group 13*
adi v1.10410 F delta-AITX-Ucs1a
adi v1.15074 F Venom dipeptidyl peptidase 4 OS Vespula vulgaris
Group 2
adi v1.20292 T Venom dipeptidyl peptidase 4 OS Vespula vulgaris
adi v1.19445 F Snake venom metalloprotease OS Philodryas olfersii
adi v1.09855 T SE-cephalotoxin OS Sepia esculenta
adi v1.16469 F L-amino-acid oxidase OS Ophiophagus hannah
adi v1.10508 T Putative protein-glutamate O-methyltransferase OS Pimpla hypochondriaca GN vpr2
adi v1.18989 F Natterin-4 OS Thalassophryne nattereri
adi v1.12311 F Toxin CfTX-1 OS Chironex fleckeri
adi v1.19322 F Venom allergen 3 OS Solenopsis richteri
adi v1.01102 T Calglandulin OS Tropidechis carinatus
Group 12*
adi v1.03437 F Calglandulin OS Tropidechis carinatus
adi v1.16440 F delta-AITX-Aas1a
Group 8
adi v1.24162 F Equinatoxin-2 OS Actinia equina
adi v1.05162 T Reticulocalbin-2 OS Crotalus adamanteus
adi v1.14946 F Snake venom 5-nucleotidase OS Gloydius brevicaudus
adi v1.06850 F Millepora cytotoxin-1 OS Millepora dichotoma
adi v1.18628 F Conodipine-M alpha chain OS Conus magus
adi v1.07444 F Astacin-like metalloprotease toxin 3 OS Loxosceles intermedia
adi v1.13648 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.15751 F Astacin-like metalloprotease toxin 1 OS Loxosceles intermedia
Group 1
adi v1.17845 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.20368 T Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.11797 F Neutral phospholipase A2 homolog taipoxin beta chain 1 OS Oxyuranus scutellatus scutellatus
adi v1.16921 F Phospholipase A2 OS Condylactis gigantea
adi v1.22096 F Ryncolin-4 OS Cerberus rynchops
adi v1.11218 F Kunitz-type serine protease inhibitor mulgin-3 OS Pseudechis australis
adi v1.23374 T Kunitz-type serine protease inhibitor Hg1 OS Hadrurus gertschi
adi v1.09427 F delta-TATX-Avl2a
adi v1.14353 T delta-ALTX-Pse2b
Group 7
adi v1.16619 F delta-TATX-Avl2a
adi v1.01810 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.04835 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.23821 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.05180 T Venom carboxylesterase-6 OS Apis mellifera
adi v1.05505 F Venom serine carboxypeptidase OS Apis mellifera
adi v1.12125 F Venom factor OS Crotalus adamanteus
Figure 3: Maximum likelihood based clustering of coral toxins. Sequence names are as following:
coral sequence id, followed by evidence for expression (T stands for True and indicates protein was
detected in proteomic analysis of nematocyst, while F stands for False and lack of detection). Last
part of sequence name is assigned annotation based on Uniprot ToxProt toxins enriched by
Group 9
Group 5
Anemone toxins. HMM-clustering generated groups are marked on the tree and groups not
generated by HMM clustering, but detected by ML clustering are marked by *.
Maximium Parsimony (MP) clustering:
adi v1.00020 F Putative endothelial lipase OS Crotalus adamanteus
Group 10
adi v1.09322 F Putative endothelial lipase OS Crotalus adamanteus
adi v1.12434 F Phospholipase A1 OS Polybia paulista
adi v1.02125 T Venom phosphodiesterase 2 OS Crotalus adamanteus
adi v1.09427 F delta-TATX-Avl2a
adi v1.14353 T delta-ALTX-Pse2b
Group 6
adi v1.16619 F delta-TATX-Avl2a
adi v1.05505 F Venom serine carboxypeptidase OS Apis mellifera
adi v1.08904 T Blarina toxin OS Blarina brevicauda GN BTX
Group 11-1
adi v1.15821 F Coagulation factor X OS Tropidechis carinatus GN
adi v1.12125 F Venom factor OS Crotalus adamanteus
adi v1.16469 F L-amino-acid oxidase OS Ophiophagus hannah
adi v1.09855 T SE-cephalotoxin OS Sepia esculenta
adi v1.19322 F Venom allergen 3 OS Solenopsis richteri
adi v1.05180 T Venom carboxylesterase-6 OS Apis mellifera
adi v1.09733 T Venom peptide isomerase heavy chain OS Agelenopsis aperta
adi v1.23604 T Astacin-like metalloprotease toxin 2 OS Loxosceles intermedia
adi v1.08969 T Phospholipase-B 81 OS Drysdalia coronoides
Group 4
adi v1.12172 T Phospholipase-B 81 OS Drysdalia coronoides
adi v1.09601 T Equinatoxin-5 OS Actinia equina
Group 13*
adi v1.10410 F delta-AITX-Ucs1a
adi v1.18989 F Natterin-4 OS Thalassophryne nattereri
adi v1.01102 T Calglandulin OS Tropidechis carinatus
Group 12*
adi v1.03437 F Calglandulin OS Tropidechis carinatus
adi v1.05162 T Reticulocalbin-2 OS Crotalus adamanteus
adi v1.16440 F delta-AITX-Aas1a
Group 8
adi v1.24162 F Equinatoxin-2 OS Actinia equina
adi v1.19802 F C-type lectin lectoxin-Thr1 OS Thrasops jacksonii
adi v1.01810 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.04835 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
Group 9
adi v1.23821 F Putative lysosomal acid lipase/cholesteryl ester hydrolase OS Crotalus adamanteus
adi v1.06850 F Millepora cytotoxin-1 OS Millepora dichotoma
adi v1.18628 F Conodipine-M alpha chain OS Conus magus
adi v1.11797 F Neutral phospholipase A2 homolog taipoxin beta chain 1 OS Oxyuranus scutellatus scutellatus
adi v1.16921 F Phospholipase A2 OS Condylactis gigantea
adi v1.15074 F Venom dipeptidyl peptidase 4 OS Vespula vulgaris
Group 2
adi v1.20292 T Venom dipeptidyl peptidase 4 OS Vespula vulgaris
adi v1.14946 F Snake venom 5-nucleotidase OS Gloydius brevicaudus
adi v1.01092 T Translationally-controlled tumor protein homolog OS Crotalus adamanteus
adi v1.10508 T Putative protein-glutamate O-methyltransferase OS Pimpla hypochondriaca GN vpr2
adi v1.12298 T Snaclec 7 OS Daboia siamensis
adi v1.12311 F Toxin CfTX-1 OS Chironex fleckeri
adi v1.06821 F Hyaluronidase-1 OS Bitis arietans
adi v1.23657 F Venom peptide isomerase heavy chain OS Agelenopsis aperta
adi v1.16374 T Venom prothrombin activator notecarin-D2 OS Notechis scutatus scutatus
adi v1.16452 F Venom phosphodiesterase 2 OS Crotalus adamanteus
adi v1.19445 F Snake venom metalloprotease OS Philodryas olfersii
adi v1.22096 F Ryncolin-4 OS Cerberus rynchops
adi v1.07444 F Astacin-like metalloprotease toxin 3 OS Loxosceles intermedia
adi v1.17845 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.15751 F Astacin-like metalloprotease toxin 1 OS Loxosceles intermedia
Group 1
adi v1.13648 F Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.20368 T Nematocyte expressed protein 6 OS Nematostella vectensis
adi v1.11218 F Kunitz-type serine protease inhibitor mulgin-3 OS Pseudechis australis
adi v1.23374 T Kunitz-type serine protease inhibitor Hg1 OS Hadrurus gertschi
Figure 4: Maximum parsimony based clustering of coral toxins.
Group 7
Group 5