posterior probabilities Values below branches: Maximum Likelihood bootstrap values.
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1 Supplementary Figure 1: Squamate molecular phylogeny. Values above branches: Bayesian posterior probabilities Values below branches: Maximum Likelihood bootstrap values.
2 Supplementary Figure 2: Bayesian analysis of representative Cystatin-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
3 Supplementary Figure 3: Bayesian analysis of representative Crotamine-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study. UPP and LOW respectively refer to transcripts from the maxillary and mandibular glands of Pogona barbata.
4 Supplementary Figure 4: Bayesian analysis of representative CRISP-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
5 Supplementary Figure 5: Bayesian analysis of representative Cobra Venom Factor-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
6 Supplementary Figure 6: Bayesian analysis of representative Kallkrein-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
7 Supplementary Figure 7: Bayesian analysis of representative AVIT-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
8 Supplementary Figure 8: Bayesian analysis of representative NGF-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
9 Supplementary Figure 9: Bayesian analysis of representative Vespryn-related sequences. In order to minimize confusion, all sequences are referred to by their SWISS-PROT accession numbers ( * Designates transcripts obtained in this study.
10 A) B) 1. GNP EKRLQPEGSC++QKMD++ B) 1. GNP EKRLQPEGSCFGQKMDRIGHVSGMGCNKFDPNKGSSSTGKK 2. TNP-c SDSKIGNGCFGFPLDRIGSVSGLGCNRIMQNPPKKFSGE- 3. DNP EVKYDPCFGHKIDRINHVSNLGCPSLRDPRPNAPSTSA 4. Lebetin -GDNKPPKKGPP-NGCFGHKIDRIGSHSGLGCNKVDDNKG BNP Rat IQERLRNSKMAHSSSCFGQKIDRIGAVSRLGCDGLRLF Supplementary Figure 10: A) Bayesian analysis of representative full-length natriuretic toxins and related sequences. In order to minimize confusion, all sequences are referred to by their SWISS- PROT accession numbers ( * Designates transcripts obtained in this study. B) ) Sequence comparison of GNP-1 (DQ139927) from V. varius (Lace Monitor), 2) TNP-c (P83230) from Oxyuranus microlepidotus (Inland taipan), 3) DNP (P28374) from Dendroaspis angusticeps (Eastern Green Mamba), 4) Lebetin (Q7LZ09) from Macrovipera lebetina (Elephant snake) and 5) BNP from the rat (P13205) brain and atria. + designates conserved functional residues.
11 Supplementary Figure 11: Liquid Chromatography/Mass Spectrometry analysis of crude mandibular secretions from Varanus varius (Lace Monitor). Reconstructed masses in Daltons are shown above each peak.
12 A) W G G HD 1*^ GAFTMPGTVWCGAGDSASDFSDLGAFQGPDSCCQQHDQCSVQITALQRKHGIFNLRPYTISHCDCDTRFRTCLMDLNDTIAD-SW 2^ GAFIMPGTLWCGAGNAASDYSQLGTEKDTDMCCRDHDHCSDTMAALEYKHGMRNYRPHTVSHCDCDNQFRSCLMNVKDRTADFSQ 3^ GAFIMPGTLWCGAGNAASDYSQLGTEKDTDMCCRDHDHCENWISALEYKHGMRNYYPSTISHCDCDNQFRSCLMKLKDGTADF-W B) H D 1* ITGGQECNEDSHPWLVLLYAEASFMCGATLLNQDWVLTAAHCYDSRPIHLYFGIHNTKQPRGHEQARDAVSTFCYPDSPGTTNSSCPSFRL DRGDDI-MLIKLNASVTYNEHIAPMAL 2* ITGGEECNEDSHPWLVVIYTEANTMCGATLLNQDWVLTAAHCYKRGKIWLNFGVHNREQARGDEEVREAVGTFCYPDSPGTTTSSCPCYTL DHGDDI-MLIKLNASVTYNEHIAPMAL 3 IIGGQECDETGHPWLALLHRSEGSTWSGVLLNRDWILTAAHCEELGPMKICFGMKNRNVLRGDEQVKVAAVKKCYPATAG-TIYNCNYVNTVLMNNDLLKRELFPMLIKLDSSVDYNERVAPLSL 4 VIGGDECDINEHPFLAFMYYSPQYFCGMTLINQEWVLTAAHCD-KTYMRIYLGIHTRSVANDDEVIRYPKEKFICPNKKKNVITD KDI-MLIRLNRPVKNSTHIAPISL 5*^ IIGGLECNQNEHRSLVLLYNSGGFFCSGTLINHEWVLTAAHCN-RENIQIKLGVHNIHVPNEDEQIRVPKEKVCCLGTMNCTQWN QDI-MLIRLNSSVNYSTHIAPLSL 6 IIGGFECNPSEHRSLVYLYNSAGFFCSGTLLNHEWVLTAAHCN-REDIQIRLGVHNVHVHYEDEQIRVPKEKLCCLSTNNCTQFS QDI-MLIRLNSPVNYSEHIAPLSL 7 IIKGFECKPHSQPWQAALFEKTRLLCGATLIAPRWLLTAAHCLKPRYIV-HLGQHNLQKEEGCEQTRTATESFPHPGFNNSL----P NKDHRNDI-MLVKMASPVSITWAVRPLTL S 1* PDRAAPLGTECDIIGWGETELTIGS----VSHIPFCASINTMDNHFCQDVSSVTITD-DMICAGVLEGGPDACKGDSGGPLLCGGQLQGLVSFGGYPCGQPMMPGVYTKIFSYREWIYSHIR---- 2* PDHAVPLGTECDIIGWGETELIVD----TASDVPLCASISTRENHFCEDAYSANMTD-DMICAGDLKGGPDACR-DSGGPLLCGGQLQGLVSFGGIPCGQPKKPGVYTKIFNYRKWIRSS PTSPASLGAECSVLGWGTTTPDDVT----LPDVPVCVNIEIFNNAVCQVAR-DLWKFTNKLCAGVDFGGKDSCK-DSGGPLVCDNQLTGNVSWGFN-CEQGEKYGYIKLIKFNF-WIQNIIQGGTG 4 PSNPPSVGSVCRIMGWGAITTSEDT----YPDVPHCANINLFNNTVCREAYNGLPAK--TLCAGVLQGGIDTCG-DSGGPLICNGQFQGILSWGSDPCAEPRKPAFYTKVFDYLPWIQSIIAGNKG 5 PSNPPSVGSVCRVMGWGTITSPEVT----YPKVPHCVNIQILHKELCEAAYPILLGNSNILCAGKLLGDKDSCK-DSGGPLICNGQIQGIVSWGGFPCAQFLSLASTPRSLIILTGSRTLW----G 6 PSNPPSMGSVCCVMGWGTITSPEVT----YPEVPHCVDINILHIPVCQAAYPTMSGK-NILCAGILEGGKDSCK-DSGGPLICNGQIQGIVSWGRFPCAQFLEPGIYTKVFDYKDWIEGIIAGNSG 7 SSRCVTAGTSCLISGWGSTSS----PQLRLPHTLRCANITIIEHQKCENAYPGNITD-TMVCASVQEGGKDSCQ-DSGGPLVCNQSLQGIISWGQDPCAITRKPGVYTKVCKYVDWIQETMKNN-G 1* * TCP- 4 TATCPP 5*^ NVICP Supplemenary Figure 12: A) Alignment of Type III PLA 2 : 1. Varanus varius (Lace Monitor) DQ and Heloderma suspectum (Gila Monster) 2. P80003 and 3. P B) Alignment of representative kallikrein (minus signal peptides): toxin forms 1. Varanus mitchelli (Mitchell s Water Monitor) DQ139915, 2. Varanus acanthurus (Spiny-tailed Monitor) DQ139914, 3. Heloderma horridum (Beaded Lizard) P43685, 4. Bothrops jararaca (Jararaca) P81661, 5. Philodryas olfersii (Argentine Racer) DQ139923, 6. Lapemis curtus (Spine-bellied Sea Snake) Q5MCS0 and tissue kallikrein 7. Homo sapiens (Human) Q9UBX7. * Designates transcripts obtained in this study. ^ Designates partial sequence.
13 KR EXXF 1* MIVFILL-SLAAVLQQFVA-D--VNFESESPRRTEKQTEIVDMHNSFRRSVNPTARNMLKMEWYPEAADNAERWAYQCI-YDHSANSERVIGGIQCGENIYKSSNPRAWTEIIQSWYDEIQNFEY 2 MIAFSLL-CFAAVLQQSFG-N--VDFNSESTRRKKKQKEIVDLHNSLRRRVSPTASNMLKMEWYPEAASNAERWANTCS-LNHSPDNLRVLEGIQCGESIYMSSNARTWTEIIHLWHDEYKNFVY 3* MFVFILL-SLAAVLQQSFG-N--VDFNSESPRIKAKQREIVDKHNAFRRSVRPTASNMLRMEWYSEAASNAERWAYRCI-LDHSPKTSRILNGIKCGENIYMSSIPMTWIDIIKLWHDEYKNFIY 4* MIVFILL-SLAAVLEQSFG-N--VDFNSESPRRPEKQKEIVDRHNSFRRSVRPTASNMLKMEWYSEAASNAERWAYRCN-LGHSPDSSRILDGIKCGENIYMSSNPRAWTEILQLWYDEYKNFVY 5 MIAFIVLLSLAAVLQQSSG-T--ADFASESSNKKNYQKEIVDKHNALRRSVKPTARNMLQMKWNSRAAQNAKRWANRCT-FAHSPPNKRTVGKLRCGENIFMSSQPFPWSGVVQAWYDEIKNFVY 6* MIAFIVLLSLAAVLQQSSG-T--VDFASESSNKKDYRKEIVDKHNDLRRSVKPTARNMLQMKWNSRAAQNAKRWANRCT-FAHSPPYTRTVGKLRCGENIFMSSQPFAWSGVVQAWYDEVKKFVY 7* MILLKLYLTLAAILCQSRGMTS-LDLDDLMTTNPEIQNEIINKHNDLRRTVDPPAKNMLKMSWDNIIAESAKRAALRCNYKEHTSIAERTIGGVVCGENHFMSSNPRTWSSSIQSWFDERNSFMF 8 MILLSLYLCLAAMLHQSEGEASPK-LPGLMTSNPDQQTEITDKHNNLRRIVEPTASNMLKMTWSNKIAQNAQRSANQCT-LEHTSKEERTIDGVECGENLFFSSAPYTWSYAIQNWFDERKYFRF 1* GVGANPPGSVIGHYTQIVWYKSYRIGCAAAYCP-SYPYNYFYVCQYCPTGNMEGLTATPYTSGPTCADCPSHCDDGLCTNPCPITNTFTNCDSLLQQNSCED--SYIKTNCGASCFCQDKII 2 GVGANPPGSVTGHYTQIVWYQTYRAGCAVSYCP-SSAWSYFYVCQYCPSGNFQGKTATPYKLGPPCGDCPSACDNGLCTNPCTIYNKLTNCDSLLKQSSCQD--DWIKSNCPASCFCRNKII 3* GVGANPPGSVIGHYTQIVWYKSYRVGCAASYCP-SSSYNYFYVCQYCPAGNFAGLTATPYKSGPTCGDCPSACDNGLCTNPCSREDVFMNCKSLVAQSNCQD--DYIRKNCPATCFCPNK-- 4* GVGANPPGSVTGHFSQMVWYKSYRIGCAAAYCP-SSGYSYFYVCQYCPIGNIEGSTATPYKSGPTCGDCPSACDNGLCTNPCLREDKFTNCKSLVQQNSCQH--DWTRKNCPATCFCHN--- 5 GIGAKPPGSVIGHYTQVVWYKSYLIGCASAKCS-SS--KYLYVCQYCPAGNIRGSIATPYKSGPPCADCPSACVNKLCTNPCKRNNDFSNCKSLAKKSKCQT--EWIKKKCPASCFCHNKII 6* GIGAKPPSSVIGHYTQVVWYKSHLLGCASAKCS-ST--KYLYVCQYCPAGNIIGSIATPYKSGPPCGDCPSACDNGLCTNPCKHNNDFSNCKALAKKSKCQT--EWIKSKCPATCFCRTEII 7* GFGPTIPGVMVGHYTQVVWYKSYKVGCAINLCPAQSL-KYFQVCQYCPGGNVAGRKYEPYTIGEPCAACPKDCDNGLCTNPCAYNDDYTSCPDLTKQVGCNHPV---TANCKASCQCTTEIQ 8 NYGPTAQNVMIGHYTQVVWYRSYELGCAIAYCPDQPTYKYYQVCQYCPGGNIRSRKYTPYSIGPPCGDCPDACDNGLCTNPCKQNDVYNNCPDLKKQVGCGH--P-IMKDCMATCKCLTEIK Supplementary Figure 13: Sequence comparison of CRISP toxins: 1) DQ Enhydris polylepis (Macleay s water snake), 2) Q7T1K6 Naja atra (Taiwanese cobra), 3) DQ Dispholidus typus (Boomslang), 4) DQ Trimorphodon biscutatus (Lyre snake), 5) Q8AVA4 Pseudechis australis (Mulga snake), 6) DQ Oxyuranus microlepidotus (Inland taipan), 7) DQ Varanus varius (Lace monitor) and 8) Q91055 Heloderma horridum (Beaded lizard).
Supporting Information
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