{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T11:56:35Z","timestamp":1793706995413,"version":"4.6.1"},"reference-count":60,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2021,6,29]],"date-time":"2021-06-29T00:00:00Z","timestamp":1624924800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005278","name":"Universidad de Antioquia","doi-asserted-by":"publisher","award":["Sostenibilidad"],"award-info":[{"award-number":["Sostenibilidad"]}],"id":[{"id":"10.13039\/501100005278","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019867","name":"Universidad Cooperativa de \u200bColo\u200bmb\u200bia","doi-asserted-by":"publisher","award":["INFETARE"],"award-info":[{"award-number":["INFETARE"]}],"id":[{"id":"10.13039\/501100019867","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Nowadays, spider venom research focuses on the neurotoxic activity of small peptides. In this study, we investigated high-molecular-mass compounds that have either enzymatic activity or housekeeping functions present in either the venom gland or venom of Pamphobeteus verdolaga. We used proteomic and transcriptomic-assisted approaches to recognize the proteins sequences related to high-molecular-mass compounds present in either venom gland or venom. We report the amino acid sequences (partial or complete) of 45 high-molecular-mass compounds detected by transcriptomics showing similarity to other proteins with either enzymatic activity (i.e., phospholipases A2, kunitz-type, hyaluronidases, and sphingomyelinase D) or housekeeping functions involved in the signaling process, glucanotransferase function, and beta-N-acetylglucosaminidase activity. MS\/MS analysis showed fragments exhibiting a resemblance similarity with different sequences detected by transcriptomics corresponding to sphingomyelinase D, hyaluronidase, lycotoxins, cysteine-rich secretory proteins, and kunitz-type serine protease inhibitors, among others. Additionally, we report a probably new protein sequence corresponding to the lycotoxin family detected by transcriptomics. The phylogeny analysis suggested that P. verdolaga includes a basal protein that underwent a duplication event that gave origin to the lycotoxin proteins reported for Lycosa sp. This approach allows proposing an evolutionary relationship of high-molecular-mass proteins among P. verdolaga and other spider species.<\/jats:p>","DOI":"10.3390\/toxins13070453","type":"journal-article","created":{"date-parts":[[2021,6,29]],"date-time":"2021-06-29T22:39:43Z","timestamp":1625006383000},"page":"453","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Analysis of High Molecular Mass Compounds from the Spider Pamphobeteus verdolaga Venom Gland. A Transcriptomic and MS ID Approach"],"prefix":"10.3390","volume":"13","author":[{"given":"Sebastian","family":"Estrada-G\u00f3mez","sequence":"first","affiliation":[{"name":"Grupo de Toxinolog\u00eda\u2014Serpentario, Facultad de Ciencias Farmac\u00e9uticas y Alimentarias, Universidad de Antioquia UdeA, Carrera 53 No. 61-30, Medell\u00edn 050010, Colombia"},{"name":"Facultad de Ciencias Farmac\u00e9uticas y Alimentarias, Universidad de Antioquia UdeA, Calle 70 No. 52-21, Medell\u00edn 050010, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1332-5106","authenticated-orcid":false,"given":"Leidy Johana","family":"Vargas-Mu\u00f1oz","sequence":"additional","affiliation":[{"name":"Facultad de Medicina, Universidad Cooperativa de Colombia, Calle 50 A No. 41-20, Medell\u00edn 050012, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9979-8416","authenticated-orcid":false,"given":"Cesar","family":"Segura Latorre","sequence":"additional","affiliation":[{"name":"Grupo de Toxinolog\u00eda\u2014Serpentario, Facultad de Ciencias Farmac\u00e9uticas y Alimentarias, Universidad de Antioquia UdeA, Carrera 53 No. 61-30, Medell\u00edn 050010, Colombia"},{"name":"Unidad de Espectrometr\u00eda de Masas, Sede de Investigaci\u00f3n Universitaria, Universidad de Antioquia UdeA, Carrera 53 No. 61-30, Medell\u00edn 050010, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4768-4919","authenticated-orcid":false,"given":"Monica Maria","family":"Saldarriaga-Cordoba","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n en Recursos Naturales y Sustentabilidad, Universidad Bernardo O\u2019Higgins, Avenida Viel 1497, Santiago 7750000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7905-5052","authenticated-orcid":false,"given":"Claudia Marcela","family":"Arenas-G\u00f3mez","sequence":"additional","affiliation":[{"name":"Grupo de G\u00e9netica, Regeneraci\u00f3n y C\u00e1ncer, Universidad de Antioquia UdeA, Carrera 53 No. 61-30, Medell\u00edn 050010, Colombia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1146\/annurev-ento-120811-153650","article-title":"Spider-venom peptides: Structure, pharmacology, and potential for control of insect pests","volume":"58","author":"King","year":"2013","journal-title":"Annu. Rev. Entomol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1111\/1744-7917.12305","article-title":"Identification and characterization of toxins in the venom gland of the Chinese bird spider, Haplopelma hainanum, by transcriptomic analysis","volume":"23","author":"Cheng","year":"2016","journal-title":"Insect Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1016\/j.toxicon.2008.03.024","article-title":"Molecular diversification based on analysis of expressed sequence tags from the venom glands of the Chinese bird spider Ornithoctonus huwena","volume":"51","author":"Jiang","year":"2008","journal-title":"Toxicon"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2792","DOI":"10.1021\/pr0700192","article-title":"Proteomic and peptidomic characterization of the venom from the Chinese bird spider, Ornithoctonus huwena Wang","volume":"6","author":"Yuan","year":"2007","journal-title":"J. Proteome Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jprot.2016.01.013","article-title":"Venomous extract protein profile of Brazilian tarantula Grammostola iheringi: Searching for potential biotechnological applications","volume":"136","author":"Borges","year":"2016","journal-title":"J. Proteom."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1892","DOI":"10.1002\/pmic.200600785","article-title":"Proteomic and peptidomic analysis of the venom from Chinese tarantula Chilobrachys jingzhao","volume":"7","author":"Liao","year":"2007","journal-title":"Proteomics"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cifuentes, Y., Estrada-Gomez, S., Vargas Munoz, L.J., and Perafan, C. (2016). Description and molecular characterization of a new species of tarantula, Pamphobeteus verdolaga, from Colombia (Aranae: Mygalomorphae: Theraphosidae). Zoologia, 33.","DOI":"10.1590\/s1984-4689zool-20160113"},{"key":"ref_8","first-page":"301","article-title":"Extraction and partial characterization of venom from the Colombian spider Pamphobeteus aff. nigricolor (Aranae:Theraphosidae)","volume":"76C","year":"2013","journal-title":"Toxicon"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.toxicon.2017.02.004","article-title":"Identifying different transcribed proteins in the newly described Theraphosidae Pamphobeteus verdolaga","volume":"129","author":"Cifuentes","year":"2017","journal-title":"Toxicon"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Estrada-Gomez, S., Caldas Cardoso, F., Vargas-Munoz, L.J., Quintana-Castillo, J.C., Arenas Gomez, C.M., and Pineda, S.S. (2019). Venomic, transcriptomic, and bioactivity analyses of Pamphobeteus verdolaga venom reveal complex disulfide-rich peptides that modulate calcium channels. Toxins, 11.","DOI":"10.3390\/toxins11090496"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1223","DOI":"10.1016\/j.toxicon.2012.08.018","article-title":"Identification, cDNA cloning and heterologous expression of a hyaluronidase from the tarantula Brachypelma vagans venom","volume":"60","author":"Clement","year":"2012","journal-title":"Toxicon"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.toxicon.2014.11.233","article-title":"Scorpion venom components as potential candidates for drug development","volume":"93","author":"Ortiz","year":"2015","journal-title":"Toxicon"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"15","DOI":"10.2174\/1570193X1101140402100416","article-title":"Purification and Characterization of Venom Components as Source for Antibiotics","volume":"11","year":"2014","journal-title":"Mini-Rev. Org. Chem."},{"key":"ref_14","unstructured":"Vargas Munoz, L.J., Estrada-Gomez, S., and Escobar, J. (2015). Snake and scorpion toxins venoms, a natural source of molecules with antimicrobial activity. Curare, 2."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1016\/j.toxicon.2005.02.018","article-title":"Structure-function relationships and mechanism of anticoagulant phospholipase A2 enzymes from snake venoms","volume":"45","author":"Kini","year":"2005","journal-title":"Toxicon"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"15307","DOI":"10.1021\/bi00046a040","article-title":"Phospholipase A2 engineering. The roles of disulfide bonds in structure, conformational stability, and catalytic function","volume":"34","author":"Zhu","year":"1995","journal-title":"Biochemistry"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"18321","DOI":"10.1074\/jbc.M008837200","article-title":"A novel group of phospholipase A2s preferentially expressed in type 2 helper T cells","volume":"276","author":"Ho","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1093\/gbe\/evv253","article-title":"Effects of Gene Duplication, Positive Selection, and Shifts in Gene Expression on the Evolution of the Venom Gland Transcriptome in Widow Spiders","volume":"8","author":"Haney","year":"2016","journal-title":"Genome Biol. Evol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.toxicon.2016.08.002","article-title":"Description of Loxtox protein family and identification of a new group of Phospholipases D from Loxosceles similis venom gland","volume":"120","author":"Dantas","year":"2016","journal-title":"Toxicon"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2529","DOI":"10.1002\/jcb.23177","article-title":"The relationship between calcium and the metabolism of plasma membrane phospholipids in hemolysis induced by brown spider venom phospholipase-D toxin","volume":"112","author":"Souza","year":"2011","journal-title":"J. Cell. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"101070","DOI":"10.1016\/j.plipres.2020.101070","article-title":"Structural insights into phospholipase D function","volume":"81","author":"Yao","year":"2021","journal-title":"Prog. Lipid Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/S0090-6980(02)00020-5","article-title":"Phospholipase A2 enzymes","volume":"68\u201369","author":"Kudo","year":"2002","journal-title":"Prostaglandins Other Lipid Mediat."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.biochi.2011.08.003","article-title":"Identification and characterisation of Kunitz-type plasma kallikrein inhibitors unique to Oxyuranus sp. snake venoms","volume":"94","author":"Earl","year":"2012","journal-title":"Biochimie"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.ijbiomac.2014.02.058","article-title":"Characterization of a Kunitz-type protease inhibitor peptide (Rusvikunin) purified from Daboia russelii russelii venom","volume":"67","author":"Mukherjee","year":"2014","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.toxicon.2012.11.004","article-title":"Molecular cloning and antifibrinolytic activity of a serine protease inhibitor from bumblebee (Bombus terrestris) venom","volume":"63","author":"Qiu","year":"2013","journal-title":"Toxicon"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1006\/bbrc.2001.4878","article-title":"Purification and Characterization of a Chymotrypsin Inhibitor from the Venom of Ophiophagus hannah (King Cobra)","volume":"283","author":"Chang","year":"2001","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.cbpb.2009.02.010","article-title":"A novel serine protease inhibitor from the venom of Vespa bicolor Fabricius","volume":"153","author":"Yang","year":"2009","journal-title":"Comp. Biochem. Physiol. Part B Biochem. Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wan, H., Lee, K.S., Kim, B.Y., Zou, F.M., Yoon, H.J., Je, Y.H., Li, J., and Jin, B.R. (2013). A spider-derived Kunitz-type serine protease inhibitor that acts as a plasmin inhibitor and an elastase inhibitor. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0053343"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2550","DOI":"10.1021\/pr1000016","article-title":"Molecular diversification of peptide toxins from the tarantula Haplopelma hainanum (Ornithoctonus hainana) venom based on transcriptomic, peptidomic, and genomic analyses","volume":"9","author":"Tang","year":"2010","journal-title":"J. Proteome Res."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Yuan, C.H., He, Q.Y., Peng, K., Diao, J.B., Jiang, L.P., Tang, X., and Liang, S.P. (2008). Discovery of a distinct superfamily of Kunitz-type toxin (KTT) from tarantulas. PLoS ONE, 3.","DOI":"10.1371\/annotation\/a7561dde-3b04-4fd9-9267-463b23cc7dd0"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2488","DOI":"10.3390\/toxins5122488","article-title":"A proteomics and transcriptomics investigation of the venom from the barychelid spider Trittame loki (brush-foot trapdoor)","volume":"5","author":"Undheim","year":"2013","journal-title":"Toxins"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S0969-2126(96)00022-6","article-title":"Structure and multiple conformations of the kunitz-type domain from human type VI collagen alpha3(VI) chain in solution","volume":"4","author":"Zweckstetter","year":"1996","journal-title":"Structure"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"He, Q., Duan, Z., Yu, Y., Liu, Z., Liu, Z., and Liang, S. (2013). The venom gland transcriptome of Latrodectus tredecimguttatus revealed by deep sequencing and cDNA library analysis. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0081357"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1812","DOI":"10.1093\/molbev\/msx116","article-title":"TimeTree: A Resource for Timelines, Timetrees, and Divergence Times","volume":"34","author":"Kumar","year":"2017","journal-title":"Mol. Biol. Evol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2059","DOI":"10.1074\/jbc.273.4.2059","article-title":"Lycotoxins, antimicrobial peptides from venom of the wolf spider Lycosa carolinensis","volume":"273","author":"Yan","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1002\/psc.1040","article-title":"Lycotoxin-1 insecticidal peptide optimized by amino acid scanning mutagenesis and expressed as a coproduct in an ethanologenic Saccharomyces cerevisiae strain","volume":"14","author":"Hughes","year":"2008","journal-title":"J. Pept. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1016\/j.toxicon.2008.08.003","article-title":"Transcriptome analysis revealed novel possible venom components and cellular processes of the tarantula Chilobrachys jingzhao venom gland","volume":"52","author":"Chen","year":"2008","journal-title":"Toxicon"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.1007\/s00018-010-0359-x","article-title":"Venom components from Citharischius crawshayi spider (Family Theraphosidae): Exploring transcriptome, venomics, and function","volume":"67","author":"Peigneur","year":"2010","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Zhang, B., Liu, Q., Yin, W., Zhang, X., Huang, Y., Luo, Y., Qiu, P., Su, X., Yu, J., and Hu, S. (2006). Transcriptome analysis of Deinagkistrodon acutus venomous gland focusing on cellular structure and functional aspects using expressed sequence tags. BMC Genom., 7.","DOI":"10.1186\/1471-2164-7-152"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1586\/14789450.5.5.731","article-title":"Proteome and peptidome profiling of spider venoms","volume":"5","author":"Liang","year":"2008","journal-title":"Expert Rev. Proteom."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.cbpb.2006.08.006","article-title":"Extraction and protein component analysis of venom from the dissected venom glands of Latrodectus tredecimguttatus","volume":"145","author":"Duan","year":"2006","journal-title":"Comp. Biochem. Physiol. B Biochem. Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/355033a0","article-title":"Protein folding in the cell","volume":"355","author":"Gething","year":"1992","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Fernandes-Pedrosa Mde, F., Junqueira-de-Azevedo Ide, L., Gon\u00e7alves-de-Andrade, R.M., Kobashi, L.S., Almeida, D.D., Ho, P.L., and Tambourgi, D.V. (2008). Transcriptome analysis of Loxosceles laeta (Araneae, Sicariidae) spider venomous gland using expressed sequence tags. BMC Genom., 9.","DOI":"10.1186\/1471-2164-9-279"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1007\/978-3-642-41588-3_11","article-title":"Ubiquitylation of voltage-gated sodium channels","volume":"221","author":"Laedermann","year":"2014","journal-title":"Handb. Exp. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Cui, Y., Zhu, Y., Lin, Y., Chen, L., Feng, Q., Wang, W., and Xiang, H. (2018). New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm. BMC Genom., 19.","DOI":"10.1186\/s12864-018-4602-4"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/1678-9199-19-5","article-title":"Comparison between two methods of scorpion venom milking in Morocco","volume":"19","author":"Oukkache","year":"2013","journal-title":"J. Venom. Anim. Toxins Incl. Trop. Dis."},{"key":"ref_47","unstructured":"World Health Organization (1981). Progress in the Characterization of Venoms and Standardization of Antivenoms, WHO Offset Publication."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.biochi.2009.12.006","article-title":"Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: Biochemical and toxicological characterization","volume":"92","author":"Fernandez","year":"2010","journal-title":"Biochimie"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"D653","DOI":"10.1093\/nar\/gkq1058","article-title":"ArachnoServer 2.0, an updated online resource for spider toxin sequences and structures","volume":"39","author":"Herzig","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"UniProt (2017). UniProt: The universal protein knowledgebase. Nucleic Acids Res., 45, D158\u2013D169.","DOI":"10.1093\/nar\/gkw1099"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"NCBI (2017). Database Resources of the National Center for Biotechnology Information. Nucleic Acids Res., 44, D7\u2013D19.","DOI":"10.1093\/nar\/gkv1290"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"W695","DOI":"10.1093\/nar\/gkq313","article-title":"A new bioinformatics analysis tools framework at EMBL-EBI","volume":"38","author":"Goujon","year":"2010","journal-title":"Nucleic Acids Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/0471250953.bi0309s53","article-title":"Finding Protein and Nucleotide Similarities with FASTA","volume":"53","author":"Pearson","year":"2016","journal-title":"Curr. Protoc. Bioinform."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1093\/molbev\/msu300","article-title":"IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies","volume":"32","author":"Nguyen","year":"2015","journal-title":"Mol. Biol. Evol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"W256","DOI":"10.1093\/nar\/gkz239","article-title":"Interactive Tree Of Life (iTOL) v4: Recent updates and new developments","volume":"47","author":"Letunic","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/j.toxicon.2008.05.020","article-title":"A rational nomenclature for naming peptide toxins from spiders and other venomous animals","volume":"52","author":"King","year":"2008","journal-title":"Toxicon"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 improves signal peptide predictions using deep neural networks","volume":"37","author":"Tsirigos","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"W177","DOI":"10.1093\/nar\/gkl266","article-title":"DISULFIND: A disulfide bonding state and cysteine connectivity prediction server","volume":"34","author":"Ceroni","year":"2006","journal-title":"Nucleic Acids Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2336","DOI":"10.1093\/bioinformatics\/bti328","article-title":"Disulfide connectivity prediction using secondary structure information and diresidue frequencies","volume":"21","author":"Clote","year":"2005","journal-title":"Bioinformatics"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"W230","DOI":"10.1093\/nar\/gki412","article-title":"DiANNA: A web server for disulfide connectivity prediction","volume":"33","author":"Clote","year":"2005","journal-title":"Nucleic Acids Res."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/7\/453\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:26:46Z","timestamp":1760164006000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/7\/453"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,29]]},"references-count":60,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2021,7]]}},"alternative-id":["toxins13070453"],"URL":"https:\/\/doi.org\/10.3390\/toxins13070453","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,29]]}}}