Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels
Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium i...
- Autores:
-
Vargas Muñoz, Leidy Johana
Vargas Muñoz, Leidy Johana
Estrada Gómez, Sebastián
Caldas Cardoso, Fernanda
Quintana Castillo, Juan Carlos
Saldarriaga Córdoba, Mónica María
Arenas Gómez, Claudia Marcela
Pineda, Sandy Steffany
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2019
- Institución:
- Universidad Cooperativa de Colombia
- Repositorio:
- Repositorio UCC
- Idioma:
- OAI Identifier:
- oai:repository.ucc.edu.co:20.500.12494/15916
- Acceso en línea:
- https://hdl.handle.net/20.500.12494/15916
- Palabra clave:
- theraphosidae
Pamphobeteus
Peptides
Disulfide-rich peptide (DRP)
Inhibitory cysteine knot (ICK)
Venomics
Transcriptome
Ion channels
- Rights
- openAccess
- License
- Atribución – No comercial – Sin Derivar
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| dc.title.spa.fl_str_mv |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| title |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| spellingShingle |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels theraphosidae Pamphobeteus Peptides Disulfide-rich peptide (DRP) Inhibitory cysteine knot (ICK) Venomics Transcriptome Ion channels |
| title_short |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| title_full |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| title_fullStr |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| title_full_unstemmed |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| title_sort |
Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels |
| dc.creator.fl_str_mv |
Vargas Muñoz, Leidy Johana Vargas Muñoz, Leidy Johana Estrada Gómez, Sebastián Caldas Cardoso, Fernanda Quintana Castillo, Juan Carlos Saldarriaga Córdoba, Mónica María Arenas Gómez, Claudia Marcela Pineda, Sandy Steffany |
| dc.contributor.author.none.fl_str_mv |
Vargas Muñoz, Leidy Johana Vargas Muñoz, Leidy Johana Estrada Gómez, Sebastián Caldas Cardoso, Fernanda Quintana Castillo, Juan Carlos Saldarriaga Córdoba, Mónica María Arenas Gómez, Claudia Marcela Pineda, Sandy Steffany |
| dc.subject.spa.fl_str_mv |
theraphosidae Pamphobeteus Peptides Disulfide-rich peptide (DRP) Inhibitory cysteine knot (ICK) Venomics Transcriptome Ion channels |
| topic |
theraphosidae Pamphobeteus Peptides Disulfide-rich peptide (DRP) Inhibitory cysteine knot (ICK) Venomics Transcriptome Ion channels |
| description |
Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium influx assays, tandem mass spectrometry analysis (tMS/MS), and venom-gland transcriptome. rp-HPLC fractions of P. verdolaga venom showed activity on CaV2.2, CaV3.2, and NaV1.7 ion channels. Active fractions contained several peptides with molecular masses ranging from 3399.4 to 3839.6 Da. The tMS/MS analysis of active fraction displaying the strongest activity to inhibit calcium channels showed sequence fragments similar to one of the translated transcripts detected in the venom-gland transcriptome. The putative peptide of this translated transcript corresponded to a toxin, here named !-theraphositoxin-Pv3a, a potential ion channel modulator toxin that is, in addition, very similar to other theraphositoxins a ecting calcium channels (i.e., !-theraphotoxin-Asp1a). Additionally, using this holistic approach, we found that P. verdolaga venom is an important source of disulfide-rich proteins expressing at least eight superfamilies. |
| publishDate |
2019 |
| dc.date.issued.none.fl_str_mv |
2019-08-27 |
| dc.date.accessioned.none.fl_str_mv |
2020-01-15T15:16:33Z |
| dc.date.available.none.fl_str_mv |
2020-01-15T15:16:33Z |
| dc.type.none.fl_str_mv |
Artículo |
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http://purl.org/coar/resource_type/c_2df8fbb1 |
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http://purl.org/coar/resource_type/c_6501 |
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http://purl.org/coar/version/c_970fb48d4fbd8a85 |
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info:eu-repo/semantics/article |
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2072-6651 |
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10.3390/toxins11090496 |
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https://hdl.handle.net/20.500.12494/15916 |
| dc.identifier.bibliographicCitation.spa.fl_str_mv |
Estrada-Gomez, S., Cardoso, F. C., Vargas-Muñoz, L. J., Quintana-Castillo, J. C., Arenas Gómez, C. M., Pineda, S. S. y Saldarriaga-Cordoba, M. M. (2019). Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels. Toxins 2019, 11, 496. Recuperado de: |
| identifier_str_mv |
2072-6651 10.3390/toxins11090496 Estrada-Gomez, S., Cardoso, F. C., Vargas-Muñoz, L. J., Quintana-Castillo, J. C., Arenas Gómez, C. M., Pineda, S. S. y Saldarriaga-Cordoba, M. M. (2019). Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels. Toxins 2019, 11, 496. Recuperado de: |
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https://hdl.handle.net/20.500.12494/15916 |
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https://www.mdpi.com/2072-6651/11/9/496 |
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Toxins |
| dc.relation.references.spa.fl_str_mv |
Escoubas, P.; Rash, L. Tarantulas: Eight-legged pharmacists and combinatorial chemists. Toxicon 2004, 43, 555–574. [CrossRef] [PubMed] King, J.B.; Gross, J.; Lovly, C.M.; Piwnica-Worms, H.; Townsend, R.R. Identification of protein phosphorylation sites within ser/thr-rich cluster domains using site-directed mutagenesis and hybrid linear quadrupole ion trap fourier transform ion cyclotron resonance mass spectrometry. Rapid Commun. Mass Spectrom. 2007, 21, 3443–3451. [CrossRef] [PubMed] Nicholson, G.M. Spider venom peptides. In Handbook of Biologically Active Peptides; Hastin, A., Ed.; Elsevier: Amsterdam, The Netherlands, 2006. Nicholson, G.M. Insect-selective spider toxins targeting voltage-gated sodium channels. Toxicon 2007, 49, 490–512. [CrossRef] [PubMed] Vassilevski, A.A.; Kozlov, S.A.; Grishin, E.V. Molecular diversity of spider venom. Biochemistry (Moscow) 2009, 74, 1505–1534. [CrossRef] [PubMed] Estrada, G.S.; Vargas, M.L.; Ramirez, A.; Quintana, C.J. Milking and partial characterization Pamphobeteus spp (aranae; theraphosidae) venom, from the colombian andean region. Toxicon 2012, 60, 231–232. Rash, L.D.; Hodgson, W.C. Pharmacology and biochemistry of spider venoms. Toxicon 2002, 40, 225–254. [CrossRef] King, G.F. Modulation of insect Ca(v) channels by peptidic spider toxins. Toxicon 2007, 49, 513–530. [CrossRef] King, G.F.; Hardy, M.C. Spider-venom peptides: Structure, pharmacology, and potential for control of insect pests. Ann. Rev. Entomol. 2013, 58, 475–496. [ Pineda, S.S.; Undheim, E.A.; Rupasinghe, D.B.; Ikonomopoulou, M.P.; King, G.F. Spider venomics: Implications for drug discovery. Future Med. Chem. 2014, 6, 1699–1714. Abreu, T.F.; Sumitomo, B.N.; Nishiyama, M.Y., Jr.; Oliveira, U.C.; Souza, G.H.; Kitano, E.S.; Zelanis, A.; Serrano, S.M.; Junqueira-de-Azevedo, I.; Silva, P.I., Jr.; et al. Peptidomics of Acanthoscurria gomesiana spider venom reveals new toxins with potential antimicrobial activity. J. Proteom. 2017, 151, 232–242. [ Chen, J.; Deng, M.; He, Q.; Meng, E.; Jiang, L.; Liao, Z.; Rong, M.; Liang, S. Molecular diversity and evolution of cystine knot toxins of the tarantula Chilobrachys jingzhao. Cell. Mol. Life Sci. 2008, 65, 2431–2444. Chen, J.; Zhao, L.; Jiang, L.; Meng, E.; Zhang, Y.; Xiong, X.; Liang, S. Transcriptome analysis revealed novel possible venom components and cellular processes of the tarantula Chilobrachys jingzhao venom gland. Toxicon 2008, 52, 794–806. Cheng, T.C.; Long, R.W.; Wu, Y.Q.; Guo, Y.B.; Liu, D.L.; Peng, L.; Li, D.Q.; Yang, D.W.; Xu, X.; Liu, F.X.; et al. Identification and characterization of toxins in the venom gland of the chinese bird spider, Haplopelma hainanum, by transcriptomic analysis. Insect Sci. 2016, 23, 487–499. Cardoso, F.C.; Dekan, Z.; Rosengren, K.J.; Erickson, A.; Vetter, I.; Deuis, J.R.; Herzig, V.; Alewood, P.F.; King, G.F.; Lewis, R.J. Identification and characterization of protx-iii [u-trtx-tp1a], a new voltage-gated sodium channel inhibitor from venom of the tarantula Thrixopelma pruriens. Mol. Pharm. 2015, 88, 291–303. Cardoso, F.C.; Dekan, Z.; Smith, J.J.; Deuis, J.R.; Vetter, I.; Herzig, V.; Alewood, P.F.; King, G.F.; Lewis, R.J. Modulatory features of the novel spider toxin -trtx-df1a isolated from the venom of the spider Davus fasciatus. Br. J. Pharm. 2017, 174, 2528–2544 Estrada-Gomez, S.; Gomez-Rave, L.; Vargas-Munoz, L.J.; van der Meijden, A. Characterizing the biological and biochemical profile of six di erent scorpion venoms from the buthidae and scorpionidae family. Toxicon 2017, 130, 104–115 Estrada-Gomez, S.; Vargas Munoz, L.J.; Quintana Castillo, J.C. Extraction and partial characterization of venom from the colombian spider Pamphobeteus a . Nigricolor (aranae:Theraphosidae). Toxicon 2013, 76, 301–309. Cardoso, F.C.; Lewis, R.J. Sodium channels and pain: From toxins to therapies. Br. J. Pharmacol. 2018, 175, 2138–2157. Xiong, X.F.; Poulsen, M.H.; Hussein, R.A.; Norager, N.G.; Stromgaard, K. Structure-activity relationship study of spider polyamine toxins as inhibitors of ionotropic glutamate receptors. ChemMedChem 2014, 9, 2661–2670. WorldSpiderCatalog. World Spider Catalog. Bern, N.H.M., Ed.; 2019. Volume Version 20.0. Available online: http://wsc.nmbe.ch (accessed on 01 July 2019). Pineda, S.S.; Chaumeil, P.A.; Kunert, A.; Kaas, Q.; Thang, M.W.C.; Le, L.; Nuhn, M.; Herzig, V.; Saez, N.J.; Cristofori-Armstrong, B.; et al. Arachnoserver 3.0: An online resource for automated discovery, analysis and annotation of spider toxins. Bioinformatics 2018, 34, 1074–1076. Savel-Niemann, A.; Roth, D. Biochemical analysis of tarantula venom (Eurypelma californicum). Naturwissenschaften 1989, 76, 212–213. Nason, D.M.; Phillips, D.; Saccomano, N.A.; Volkmann, R.A. Calcium Channel Blocking Polypeptides from Therpahosidae Aphonopelma. U.S. Patent WO1994010196 A, 11 May 1994. Corzo, G.; Diego-Garcia, E.; Clement, H.; Peigneur, S.; Odell, G.; Tytgat, J.; Possani, L.D.; Alagon, A. An insecticidal peptide from the theraposid Brachypelma smithi spider venom reveals common molecular features among spider species from di erent genera. Peptides 2008, 29, 1901–1908. Liang, S. An overview of peptide toxins from the venom of the chinese bird spider Selenocosmia huwena wang [=Ornithoctonus huwena (wang)]. Toxicon 2004, 43, 575–585. Wilson, D.; Daly, N.L. Venomics: A mini-review. High. Throughput 2018, 7, 19. Tang, X.; Zhang, Y.; Hu,W.; Xu, D.; Tao, H.; Yang, X.; Li, Y.; Jiang, L.; Liang, S. Molecular diversification of peptide toxins from the tarantula Haplopelma hainanum (Ornithoctonus hainana) venom based on transcriptomic, peptidomic, and genomic analyses. J. Proteome Res. 2010, 9, 2550–2564 Cifuentes,Y.; Estrada-Gomez, S.; Vargas Munoz, L.J.; Perafan, C. 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Simao, F.A.; Waterhouse, R.M.; Ioannidis, P.; Kriventseva, E.V.; Zdobnov, E.M. Busco: Assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics 2015, 31, 3210–3212 Langmead, B.; Salzberg, S.L. Fast gapped-read alignment with bowtie 2. Nat. Methods 2012, 9, 357–359. Li, B.; Dewey, C.N. Rsem: Accurate transcript quantification from rna-seq data with or without a reference genome. BMC Bioinform. 2011, 12, 323. Petersen, T.N.; Brunak, S.; von Heijne, G.; Nielsen, H. Signalp 4.1: Discriminating signal peptides from transmembrane regions. Nat. Methods 2011, 8, 785–786. Goujon, M.; McWilliam, H.; Li, W.; Valentin, F.; Squizzato, S.; Paern, J.; Lopez, R. A new bioinformatics analysis tools framework at embl-ebi. Nucl. Acids Res. 2010, 38, W695–W699. King, G.F.; Gentz, M.C.; Escoubas, P.; Nicholson, G.M. A rational nomenclature for naming peptide toxins from spiders and other venomous animals. Toxicon 2008, 52, 264–276. Kearse, M.; Moir, R.; Wilson, A.; Stones-Havas, S.; Cheung, M.; Sturrock, S.; Buxton, S.; Cooper, A.; Markowitz, S.; Duran, C.; et al. Geneious basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 2012, 28, 1647–1649 Nicholas, K.B.; Nicholas, H.B.; Deerfield, D.W. Genedoc: Analysis and visualization of genetic variation. Embnew News 1997, 4, 14. Ceroni, A.; Passerini, A.; Vullo, A.; Frasconi, P. DISULFIND: A Disulfide Bonding State and Cysteine Connectivity Prediction Server. Nucleic Acids Res. 2006, 34 (Suppl. S2), W177–W181 Sairaman, A.; Cardoso, F.C.; Bispat, A.; Lewis, R.J.; Duggan, P.J.; Tuck, K.L. Synthesis and evaluation of aminobenzothiazoles as blockers of n- and t-type calcium channels. Bioorg. Med. Chem. 2018, 26, 3046–3059. |
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Vargas Muñoz, Leidy JohanaVargas Muñoz, Leidy JohanaEstrada Gómez, SebastiánCaldas Cardoso, FernandaQuintana Castillo, Juan CarlosSaldarriaga Córdoba, Mónica MaríaArenas Gómez, Claudia MarcelaPineda, Sandy Steffany112020-01-15T15:16:33Z2020-01-15T15:16:33Z2019-08-272072-665110.3390/toxins11090496https://hdl.handle.net/20.500.12494/15916Estrada-Gomez, S., Cardoso, F. C., Vargas-Muñoz, L. J., Quintana-Castillo, J. C., Arenas Gómez, C. M., Pineda, S. S. y Saldarriaga-Cordoba, M. M. (2019). Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels. Toxins 2019, 11, 496. Recuperado de:Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium influx assays, tandem mass spectrometry analysis (tMS/MS), and venom-gland transcriptome. rp-HPLC fractions of P. verdolaga venom showed activity on CaV2.2, CaV3.2, and NaV1.7 ion channels. Active fractions contained several peptides with molecular masses ranging from 3399.4 to 3839.6 Da. The tMS/MS analysis of active fraction displaying the strongest activity to inhibit calcium channels showed sequence fragments similar to one of the translated transcripts detected in the venom-gland transcriptome. The putative peptide of this translated transcript corresponded to a toxin, here named !-theraphositoxin-Pv3a, a potential ion channel modulator toxin that is, in addition, very similar to other theraphositoxins a ecting calcium channels (i.e., !-theraphotoxin-Asp1a). Additionally, using this holistic approach, we found that P. verdolaga venom is an important source of disulfide-rich proteins expressing at least eight superfamilies.https://scienti.colciencias.gov.co/cvlac/visualizador/generarCurriculoCv.do?cod_rh=0000580023https://orcid.org/0000-0003-1332-5106https://scienti.colciencias.gov.co/gruplac/jsp/visualiza/visualizagr.jsp?nro=00000000011355leidy.vargasmu@campusucc.edu.cosebastian.estrada@udea.edu.cof.caldascardoso@imb.uq.edu.aujuan.quintana@campusucc.edu.comonysalda@gmail.comclaudia.arenas@udea.edu.cosandy.spineda@gmail.com21Universidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Medicina, MedellínMedicinaMedellínhttps://www.mdpi.com/2072-6651/11/9/496ToxinsEscoubas, P.; Rash, L. Tarantulas: Eight-legged pharmacists and combinatorial chemists. Toxicon 2004, 43, 555–574. [CrossRef] [PubMed]King, J.B.; Gross, J.; Lovly, C.M.; Piwnica-Worms, H.; Townsend, R.R. 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Chem. 2018, 26, 3046–3059.theraphosidaePamphobeteusPeptidesDisulfide-rich peptide (DRP)Inhibitory cysteine knot (ICK)VenomicsTranscriptomeIon channelsVenomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium ChannelsArtículohttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1http://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionAtribución – No comercial – Sin Derivarinfo:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2PublicationTEXT2019_Estradaycolaboradores_Venomic_Transcriptomic_Bioactivity_Analyses.pdf.txt2019_Estradaycolaboradores_Venomic_Transcriptomic_Bioactivity_Analyses.pdf.txtExtracted texttext/plain91807https://repository.ucc.edu.co/bitstreams/945efac0-4dbe-4214-8a6c-40fa5552150b/download5681e258a877c9881c59e63c9af6f036MD562020_Licencia de uso_ Venomic, Transcriptomic, and 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