{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T22:18:27Z","timestamp":1777501107280,"version":"3.51.4"},"reference-count":58,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2016,12,26]],"date-time":"2016-12-26T00:00:00Z","timestamp":1482710400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Scorpion venom is a complex combinatorial library of peptides and proteins with multiple biological functions. A combination of transcriptomic and proteomic techniques has revealed its enormous molecular diversity, as identified by the presence of a large number of ion channel-targeted neurotoxins with different folds, membrane-active antimicrobial peptides, proteases, and protease inhibitors. Although the biodiversity of scorpion venom has long been known, how it arises remains unsolved. In this work, we analyzed the exon-intron structures of an array of scorpion venom protein-encoding genes and unexpectedly found that nearly all of these genes possess a phase-1 intron (one intron located between the first and second nucleotides of a codon) near the cleavage site of a signal sequence despite their mature peptides remarkably differ. This observation matches a theory of exon shuffling in the origin of new genes and suggests that recruitment of different folds into scorpion venom might be achieved via shuffling between body protein-coding genes and ancestral venom gland-specific genes that presumably contributed tissue-specific regulatory elements and secretory signal sequences.<\/jats:p>","DOI":"10.3390\/toxins9010010","type":"journal-article","created":{"date-parts":[[2016,12,28]],"date-time":"2016-12-28T11:22:14Z","timestamp":1482924134000},"page":"10","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Exon Shuffling and Origin of Scorpion Venom Biodiversity"],"prefix":"10.3390","volume":"9","author":[{"given":"Xueli","family":"Wang","sequence":"first","affiliation":[{"name":"Group of Peptide Biology and Evolution, State Key Laboratory of Integrated Management of Pest Insects &amp; Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bin","family":"Gao","sequence":"additional","affiliation":[{"name":"Group of Peptide Biology and Evolution, State Key Laboratory of Integrated Management of Pest Insects &amp; Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0614-9207","authenticated-orcid":false,"given":"Shunyi","family":"Zhu","sequence":"additional","affiliation":[{"name":"Group of Peptide Biology and Evolution, State Key Laboratory of Integrated Management of Pest Insects &amp; Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2016,12,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.jprot.2015.07.030","article-title":"Unique diversity of the venom peptides from the scorpion Androctonus bicolor revealed by transcriptomic and proteomic analysis","volume":"128","author":"Zhang","year":"2015","journal-title":"J. Proteom."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1101\/gr.3228405","article-title":"From genome to \u201cvenome\u201d: Molecular origin and evolution of the snake venom proteome inferred from phylogenetic analysis of toxin sequences and related body proteins","volume":"15","author":"Fry","year":"2005","journal-title":"Genome Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2088","DOI":"10.1093\/gbe\/evu166","article-title":"Restriction and recruitment-gene duplication and the origin and evolution of snake venom toxins","volume":"6","author":"Hargreaves","year":"2014","journal-title":"Genome Biol. Evol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1093\/molbev\/msu038","article-title":"Experimental conversion of a defensin into a neurotoxin: Implications for origin of toxic function","volume":"31","author":"Zhu","year":"2014","journal-title":"Mol. Biol. Evol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.1016\/j.bcp.2010.07.008","article-title":"Drosotoxin, a selective inhibitor of tetrodotoxin-resistant sodium channels","volume":"80","author":"Zhu","year":"2010","journal-title":"Biochem. Pharmacol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.bcp.2014.12.002","article-title":"Mesomartoxin, a new Kv1.2-selective scorpion toxin interacting with the channel selectivity filter","volume":"93","author":"Wang","year":"2015","journal-title":"Biochem. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e00047","DOI":"10.1042\/BSR20130052","article-title":"Functional evolution of scorpion venom peptides with an inhibitor cystine knot fold","volume":"33","author":"Gao","year":"2013","journal-title":"Biosci. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.biochi.2010.01.011","article-title":"Characterization of two linear cationic antimalarial peptides in the scorpion Mesobuthus eupeus","volume":"92","author":"Gao","year":"2010","journal-title":"Biochimie"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"22399","DOI":"10.1074\/jbc.M113.483875","article-title":"The X-ray crystal structure of mannose-binding lectin-associated serine proteinase-3 reveals the structural basis for enzyme inactivity associated with the Carnevale, Mingarelli, Malpuech, and Michels (3MC) syndrome","volume":"288","author":"Yongqing","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Chen, Z., Luo, F., Feng, J., Yang, W., Zeng, D., Zhao, R., Cao, Z., Liu, M., Li, W., and Jiang, L. (2013). Genomic and structural characterization of Kunitz-type peptide LmKTT-1a highlights diversity and evolution of scorpion potassium channel toxins. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0060201"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2257","DOI":"10.1007\/s00018-005-5200-6","article-title":"Phylogenetic distribution, functional epitopes and evolution of the CS\u03b1\u03b2 superfamily","volume":"62","author":"Zhu","year":"2005","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6825","DOI":"10.1016\/j.febslet.2006.11.040","article-title":"Molecular characterization of a new scorpion venom lipolysis activating peptide: Evidence for disulfide bridge-mediated functional switch of peptides","volume":"580","author":"Zhu","year":"2006","journal-title":"FEBS Lett."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2602","DOI":"10.1038\/ncomms3602","article-title":"The genome of Mesobuthus martensii reveals a unique adaptation model of arthropods","volume":"4","author":"Cao","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.toxicon.2013.07.012","article-title":"Scorpion venom components that affect ion-channels function","volume":"76","author":"Gurrola","year":"2013","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1016\/j.toxicon.2004.03.022","article-title":"Current views on scorpion toxins specific for K+-channels","volume":"43","author":"Possani","year":"2004","journal-title":"Toxicon"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1046\/j.1432-1327.1999.00625.x","article-title":"Scorpion toxins specific for Na+-channels","volume":"264","author":"Possani","year":"1999","journal-title":"Eur. J. Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"831","DOI":"10.1016\/j.toxicon.2005.09.006","article-title":"Overview of scorpion toxins specific for Na+ channels and related peptides: Biodiversity, structure-function relationships and evolution","volume":"46","author":"Possani","year":"2005","journal-title":"Toxicon"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1016\/S0165-6147(99)01398-X","article-title":"A unified nomenclature for short-chain peptides isolated from scorpion venoms: Alpha-KTx molecular subfamilies","volume":"20","author":"Tytgat","year":"1999","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/S0014-5793(98)00780-7","article-title":"Evidence for a new class of scorpion toxins active against K+ channels","volume":"431","author":"Legros","year":"1998","journal-title":"FEBS Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1671","DOI":"10.1016\/S0041-0101(98)00160-3","article-title":"Sodium channel modifiers from scorpion venom: Structure-activity relationship, mode of action and application","volume":"36","author":"Gurevitz","year":"1998","journal-title":"Toxicon"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s00239-003-2534-2","article-title":"Adaptive evolution of scorpion sodium channel toxins","volume":"58","author":"Zhu","year":"2004","journal-title":"J. Mol. Evol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/S0300-9084(00)01174-3","article-title":"Molecular mechanisms of neurotoxin action on voltage-gated sodium channels","volume":"82","author":"Catterall","year":"2000","journal-title":"Biochimie"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0896-6273(00)81133-2","article-title":"From ionic currents to molecular mechanisms: The structure and function of voltage-gated sodium channels","volume":"26","author":"Catterall","year":"2000","journal-title":"Neuron"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7363","DOI":"10.1073\/pnas.0401567101","article-title":"Multidimensional signatures in antimicrobial peptides","volume":"101","author":"Yount","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.cbpc.2015.09.002","article-title":"A single-point mutation enhances dual functionality of a scorpion toxin","volume":"179","author":"Wang","year":"2016","journal-title":"Comp. Biochem. Physiol. C Toxicol. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Diego-Garc\u00eda, E., Caliskan, F., and Tytgat, J. (2014). The Mediterranean scorpion Mesobuthus gibbosus (Scorpiones, Buthidae): Transcriptome analysis and organization of the genome encoding chlorotoxin-like peptides. BMC Genom., 15.","DOI":"10.1186\/1471-2164-15-295"},{"key":"ref_27","first-page":"239","article-title":"Origin of neurotoxins from defensins","volume":"67","author":"Zhu","year":"2015","journal-title":"Acta Physiol. Sin."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3497","DOI":"10.1529\/biophysj.107.120840","article-title":"Charged surface area of maurocalcine determines its interaction with the skeletal ryanodine receptor","volume":"95","author":"Sztretye","year":"2008","journal-title":"Biophys. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.3390\/ph3041093","article-title":"Imperatoxin A, a cell-penetrating peptide from scorpion venom, as a probe of Ca2+ release channels\/ryanodine receptors","volume":"3","author":"Gurrola","year":"2010","journal-title":"Pharmaceuticals"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"5023","DOI":"10.1046\/j.1432-1327.2000.01556.x","article-title":"Hadrurin, a new antimicrobial peptide from the venom of the scorpion Hadrurus aztecus","volume":"267","author":"Gurrola","year":"2000","journal-title":"Eur. J. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1230","DOI":"10.1096\/fj.08-122317","article-title":"Structural and functional characterization of two genetically related meucin peptides highlights evolutionary divergence and convergence in antimicrobial peptides","volume":"23","author":"Gao","year":"2009","journal-title":"FASEB J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1042\/bj3590035","article-title":"Characterization of unique amphipathic antimicrobial peptides from venom of the scorpion Pandinus imperator","volume":"359","author":"Corzo","year":"2001","journal-title":"Biochem. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3472","DOI":"10.1128\/AAC.01436-08","article-title":"Imcroporin, a new cationic antimicrobial peptide from the venom of the scorpion Isometrus maculates","volume":"53","author":"Zhao","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.toxicon.2009.10.027","article-title":"Parabutoporin, a cationic amphipathic peptide from scorpion venom: Much more than an antibiotic","volume":"55","author":"Remijsen","year":"2010","journal-title":"Toxicon"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.peptides.2013.03.026","article-title":"Three new antimicrobial peptides from the scorpion Pandinus imperator","volume":"45","author":"Zeng","year":"2013","journal-title":"Peptides"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1139\/o98-026","article-title":"Structure-function relationships of antimicrobial peptides","volume":"76","author":"Hwang","year":"1998","journal-title":"Biochem. Cell Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1016\/0041-0101(81)90077-5","article-title":"Neurotoxins, protease inhibitors and histamine releasers in the venom of the Indian red scorpion (Buthus tamulus): Isolation and partial characterization","volume":"19","author":"Chhatwal","year":"1981","journal-title":"Toxicon"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Schwartz, E.F., Diego-Garcia, E., de la Vega, R.C.R., and Possani, L.D. (2007). Transcriptome analysis of the venom gland of the Mexican scorpion Hadrurus gertschi (Arachnida: Scorpiones). BMC Genom., 8.","DOI":"10.1186\/1471-2164-8-119"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"13813","DOI":"10.1074\/jbc.M112.343996","article-title":"Hg1, novel peptide inhibitor specific for Kv1.3 channels from first scorpion Kunitz-type potassium channel toxin family","volume":"287","author":"Chen","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.toxicon.2013.05.022","article-title":"Cloning and characterization of a novel Kunitz-type inhibitor from scorpion with unique cysteine framework","volume":"72","author":"Chen","year":"2013","journal-title":"Toxicon"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Chen, Z., Wang, B., Hu, J., Yang, W., Cao, Z., Zhuo, R., Li, W., and Wu, Y. (2013). SjAPI, the first functionally characterized Ascaris-type protease inhibitor from animal venoms. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0057529"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1016\/S0969-2126(00)00067-8","article-title":"High-resolution structure of Ascaris trypsin inhibitor in solution: Direct evidence for a pH-induced conformational transition in the reactive site","volume":"2","author":"Grasberger","year":"1994","journal-title":"Structure"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Patthy, L. (2008). Introns: Phase Compatibility. eLS.","DOI":"10.1002\/9780470015902.a0005083.pub2"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.febslet.2004.08.045","article-title":"Insertion of spliceosomal introns in proto-splice sites: The case of secretory signal peptides","volume":"575","author":"Tordai","year":"2004","journal-title":"FEBS Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1016\/j.febslet.2006.01.094","article-title":"A possible role of exon-shuffling in the evolution of signal peptides of human proteins","volume":"580","author":"Vibranovski","year":"2006","journal-title":"FEBS Lett."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.1016\/j.peptides.2005.10.010","article-title":"Cloning and characterization of a novel calcium channel toxin-like gene BmCa1 from Chinese scorpion Mesobuthus martensii Karsch","volume":"27","author":"Cao","year":"2006","journal-title":"Peptides"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.1096\/fj.02-1044fje","article-title":"Evolutionary origin of inhibitor cystine knot peptides","volume":"17","author":"Zhu","year":"2003","journal-title":"FASEB J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1007\/PL00006457","article-title":"Dynamic diversification from a putative common ancestor of scorpion toxins affecting sodium, potassium, and chloride channels","volume":"48","author":"Froy","year":"1999","journal-title":"J. Mol. Evol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1002\/psc.578","article-title":"Arthropod defensins illuminate the divergence of scorpion neurotoxins","volume":"10","author":"Froy","year":"2004","journal-title":"J. Pept. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.peptides.2013.12.012","article-title":"Genomic and functional characterization of three new venom peptides from the scorpion Heterometrus spinifer","volume":"53","author":"Wu","year":"2014","journal-title":"Peptides"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"7727","DOI":"10.1073\/pnas.93.15.7727","article-title":"Exon shuffling and the origin of the mitochondrial targeting function in plant cytochrome c1 precursor","volume":"93","author":"Long","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1751","DOI":"10.1007\/s00018-004-4149-1","article-title":"The scorpine family of defensins: Gene structure, alternative polyadenylation and fold recognition","volume":"61","author":"Zhu","year":"2004","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1016\/j.tig.2003.10.010","article-title":"Arthropod and mollusk defensins\u2014Evolution by exon-shuffling","volume":"19","author":"Froy","year":"2003","journal-title":"Trends Genet."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1642","DOI":"10.1101\/gr.520702","article-title":"Signatures of domain shuffling in the human genome","volume":"12","author":"Kaessmann","year":"2002","journal-title":"Genome Res."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2456","DOI":"10.3390\/toxins5122456","article-title":"Evolution stings: The origin and diversification of scorpion toxin peptide scaffolds","volume":"5","author":"Sunagar","year":"2013","journal-title":"Toxins"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5141","DOI":"10.1096\/fj.12-218479","article-title":"A natural point mutation changes both target selectivity and mechanism of action of sea anemone toxins","volume":"26","author":"Peigneur","year":"2012","journal-title":"FASEB J."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"8906","DOI":"10.1073\/pnas.1214062110","article-title":"Multiple actions of phi-LITX-Lw1a on ryanodine receptors reveal a functional link between scorpion DDH and ICK toxins","volume":"110","author":"Smith","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1016\/j.bcp.2012.01.021","article-title":"Purification, molecular cloning and functional characterization of HelaTx1 (Heterometrus laoticus): The first member of a new kappa-KTX subfamily","volume":"83","author":"Vandendriessche","year":"2012","journal-title":"Biochem. Pharmacol."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/9\/1\/10\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:29:19Z","timestamp":1760210959000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/9\/1\/10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,12,26]]},"references-count":58,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,1]]}},"alternative-id":["toxins9010010"],"URL":"https:\/\/doi.org\/10.3390\/toxins9010010","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,12,26]]}}}