{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T05:06:27Z","timestamp":1788498387888,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2022,9,18]],"date-time":"2022-09-18T00:00:00Z","timestamp":1663459200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Foundation for Poison Control","award":["NPCC-201901"],"award-info":[{"award-number":["NPCC-201901"]}]},{"name":"Foundation for Poison Control","award":["TCVGH-1117201"],"award-info":[{"award-number":["TCVGH-1117201"]}]},{"DOI":"10.13039\/501100010101","name":"Taichung Veterans General Hospital","doi-asserted-by":"publisher","award":["NPCC-201901"],"award-info":[{"award-number":["NPCC-201901"]}],"id":[{"id":"10.13039\/501100010101","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010101","name":"Taichung Veterans General Hospital","doi-asserted-by":"publisher","award":["TCVGH-1117201"],"award-info":[{"award-number":["TCVGH-1117201"]}],"id":[{"id":"10.13039\/501100010101","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Reports of bite from Protobothrops mucrosquamatus (Pmu) are frequent in Taiwan, and its wide-spread distribution and diverse habitats drove us to investigate its envenoming effects and relevant venom variations. We used reversed-phase high-performance liquid chromatography and mass spectrometry to analyze 163 Pmu venom samples collected from northern and southeastern Taiwan. Twenty-two major protein fractions were separated and analyzed, and their contents were determined semi-quantitatively. The results showed that despite the trivial differences in the protein family, there is an existing variation in acidic phospholipases A2s, serine proteinases, metalloproteinases, C-type lectin-like proteins, and other less abundant components in the Pmu venoms. Moreover, clinical manifestations of 209 Pmu envenomed patients hospitalized in northern or southeastern Taiwan revealed significant differences in local symptoms, such as ecchymosis and blistering. The mechanism of these local effects and possibly relevant venom components were examined. Further analysis showed that certain venom components with inter-population variation might work alone or synergistically with others to aggravate the local effects. Therefore, our findings of the venom variation may help one to improve antivenom production and better understand and manage Pmu bites.<\/jats:p>","DOI":"10.3390\/toxins14090643","type":"journal-article","created":{"date-parts":[[2022,9,20]],"date-time":"2022-09-20T04:28:55Z","timestamp":1663648135000},"page":"643","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Comparison of Protein Variation in Protobothrops mucrosquamatus Venom between Northern and Southeast Taiwan and Association with Human Envenoming Effects"],"prefix":"10.3390","volume":"14","author":[{"given":"Liao-Chun","family":"Chiang","sequence":"first","affiliation":[{"name":"College of Life Sciences, National Tsing Hua University, Hsinchu City 300, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun-Yi","family":"Chien","sequence":"additional","affiliation":[{"name":"Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan County 333, Taiwan"},{"name":"Clinical Proteomics Core Laboratory, Chang Gung Memorial Hospital, Taoyuan County 333, Taiwan"},{"name":"Department of Biochemistry and Molecular Biology, College of Medicine, Chang Gung University, Taoyuan County 333, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hung-Yuan","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, E-Da Hospital, Kaohsiung County 824, Taiwan"},{"name":"The School of Chinese Medicine for Post Baccalaureate, I-Shou University, Kaohsiung County 840, Taiwan"},{"name":"Department of Safety, Health and Environmental Engineering, National Kaohsiung University of Science and Technology, Kaohsiung County 811, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yen-Chia","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, Taipei Veterans General Hospital, Taipei City 112, Taiwan"},{"name":"School of Medicine, National Yang Ming Chiao Tung University, Taipei City 112, Taiwan"},{"name":"Department of Emergency Medicine, National Defense Medical Center, Taipei City 114, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8561-0582","authenticated-orcid":false,"given":"Yan-Chiao","family":"Mao","sequence":"additional","affiliation":[{"name":"Department of Emergency Medicine, National Defense Medical Center, Taipei City 114, Taiwan"},{"name":"Division of Clinical Toxicology, Department of Emergency Medicine, Taichung Veterans General Hospital, Taichung City 407, Taiwan"},{"name":"College of Medicine, National Chung Hsing University, Taichung City 402, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen-Guey","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Life Sciences, National Tsing Hua University, Hsinchu City 300, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,18]]},"reference":[{"key":"ref_1","first-page":"111","article-title":"Protobothrops mucrosquamatus (Cantor, 1839), a highly venomous species added to the snake fauna of Thailand (Squamata: Viperidae)","volume":"17","author":"Vasaruchapong","year":"2017","journal-title":"Trop. Nat. Hist."},{"key":"ref_2","first-page":"1","article-title":"WHO Expert Committee on Biological Standardization Sixty-seventh report","volume":"1004","author":"Cichutek","year":"2017","journal-title":"Tech. Rep. Ser.-World Health Organ."},{"key":"ref_3","unstructured":"Gopalakrishnakone, P., Faiz, S.M.A., Gnanathasan, C.A., Habib, A.G., Fernando, R., and Yang, C.C. (2013). Epidemiology of Snake Envenomation in Taiwan. Toxinology: Clinical Toxinology, Springer."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1080\/15563650.2020.1762892","article-title":"Clinical manifestations and treatments of Protobothrops mucrosquamatus bite and associated factors for wound necrosis and subsequent debridement and finger or toe amputation surgery","volume":"59","author":"Mao","year":"2020","journal-title":"Clin. Toxicol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5628","DOI":"10.1016\/j.jprot.2012.08.008","article-title":"Snake venomics and antivenomics of Protobothrops mucrosquamatus and Viridovipera stejnegeri from Taiwan: Keys to understand the variable immune response in horses","volume":"75","author":"Villalta","year":"2012","journal-title":"J. Proteom."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2640","DOI":"10.1093\/gbe\/evx199","article-title":"Population Genomic Analysis of a Pitviper Reveals Microevolutionary Forces Underlying Venom Chemistry","volume":"9","author":"Aird","year":"2017","journal-title":"Genome Biol. Evol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1016\/j.tips.2020.05.006","article-title":"Causes and Consequences of Snake Venom Variation","volume":"41","author":"Casewell","year":"2020","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2576","DOI":"10.1016\/j.jprot.2012.02.035","article-title":"Venom variability and envenoming severity outcomes of the Crotalus scutulatus scutulatus (Mojave rattlesnake) from Southern Arizona","volume":"75","author":"Massey","year":"2012","journal-title":"J. Proteom."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.toxicon.2019.09.022","article-title":"Case reports of envenomation and venom composition differences between two Arizona populations of the Southwestern Speckled Rattlesnake, Crotalus pyrrhus (Cope, 1867)","volume":"171","author":"Cochran","year":"2019","journal-title":"Toxicon"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.toxicon.2017.04.002","article-title":"Geographical variability of the venoms of four populations of Bothrops asper from Panama: Toxicological analysis and neutralization by a polyvalent antivenom","volume":"132","author":"Salazar","year":"2017","journal-title":"Toxicon"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Tan, C.H., Palasuberniam, P., and Tan, K.Y. (2021). Snake venom proteomics, immunoreactivity and toxicity neutralization studies for the asiatic mountain pit vipers, Ovophis convictus, Ovophis tonkinensis, and Hime Habu, Ovophis okinavensis. Toxins, 13.","DOI":"10.3390\/toxins13080514"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"28","DOI":"10.4269\/ajtmh.2009.80.28","article-title":"Differences in clinical profiles of patients with Protobothrops mucrosquamatus and Viridovipera stejnegeri envenoming in Taiwan","volume":"80","author":"Chen","year":"2009","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1177\/102490791602300402","article-title":"Can surgery in patient with Protobothrops mucrosquamatus envenomation be predicted in emergency department?","volume":"23","author":"Su","year":"2016","journal-title":"Hong Kong J. Emerg. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1042\/bj20030818","article-title":"Venom phospholipases A2 of bamboo viper (Trimeresurus stejnegeri): Molecular characterization, geographic variations and evidence of multiple ancestries","volume":"377","author":"Tsai","year":"2004","journal-title":"Biochem. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1007\/s00239-002-2403-4","article-title":"Genetic and ecological correlates of intraspecific variation in pitviper venom composition detected using matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) and isoelectric focusing","volume":"56","author":"Creer","year":"2003","journal-title":"J. Mol. Evol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Aird, S.D., Watanabe, Y., Villar-Briones, A., Roy, M.C., Terada, K., and Mikheyev, A.S. (2013). Quantitative high-throughput profiling of snake venom gland transcriptomes and proteomes (Ovophis okinavensis and Protobothrops flavoviridis). BMC Genom., 14.","DOI":"10.1186\/1471-2164-14-790"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.bbapap.2004.08.006","article-title":"Comparative proteomics and subtyping of venom phospholipases A2 and disintegrins of Protobothrops pit vipers","volume":"1702","author":"Tsai","year":"2004","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1006\/abbi.2000.2229","article-title":"Differential expression and geographic variation of the venom phospholipases A2 of Calloselasma rhodostoma and Trimeresurus mucrosquamatus","volume":"387","author":"Tsai","year":"2001","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1081\/TXR-120026919","article-title":"Variations of phospholipases A2 in the geographic venom samples of pitvipers","volume":"22","author":"Tsai","year":"2003","journal-title":"J. Toxicol. Toxin Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"888","DOI":"10.1016\/j.toxicon.2009.12.016","article-title":"Induction of microvascular leakage and histamine release by promutoxin, an Arg49 phospholipase A2","volume":"55","author":"Wei","year":"2010","journal-title":"Toxicon"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/S0196-9781(96)00259-8","article-title":"Isolation and amino acid sequence of flavostatin, a novel disintegrin from the venom of Trimeresurus flavoviridis","volume":"18","author":"Maruyama","year":"1997","journal-title":"Peptides"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1016\/j.toxicon.2005.02.012","article-title":"Structural considerations of the snake venom metalloproteinases, key members of the M12 reprolysin family of metalloproteinases","volume":"45","author":"Fox","year":"2005","journal-title":"Toxicon"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1093\/jb\/mvg202","article-title":"A novel metalloprotease from Gloydius halys venom induces endothelial cell apoptosis through its protease and disintegrin-like domains","volume":"134","author":"You","year":"2003","journal-title":"J. Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1042\/bj3570719","article-title":"Purification, molecular cloning and mechanism of action of graminelysin I, a snake-venom-derived metalloproteinase that induces apoptosis of human endothelial cells","volume":"357","author":"Wu","year":"2001","journal-title":"Biochem. J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1002\/jcb.20322","article-title":"Characterization of events associated with apoptosis\/anoikis induced by snake venom metalloproteinase BaP1 on human endothelial cells","volume":"94","author":"Valverde","year":"2005","journal-title":"J. Cell. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Herrera, C., Escalante, T., Voisin, M.B., Rucavado, A., Moraz\u00e1n, D., Mac\u00eado, J.K., Calvete, J.J., Sanz, L., Nourshargh, S., and Guti\u00e9rrez, J.M. (2015). Tissue localization and extracellular matrix degradation by PI, PII and PIII snake venom metalloproteinases: Clues on the mechanisms of venom-induced hemorrhage. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003731"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"100052","DOI":"10.1016\/j.toxcx.2020.100052","article-title":"Snake Venom Metalloproteinases (SVMPs): A structure-function update","volume":"7","author":"Olaoba","year":"2020","journal-title":"Toxicon X"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Baldo, C., Jamora, C., Yamanouye, N., Zorn, T.M., and Moura-da-Silva, A.M. (2010). Mechanisms of vascular damage by hemorrhagic snake venom metalloproteinases: Tissue distribution and in situ hydrolysis. PLoS Negl. Trop. Dis., 4.","DOI":"10.1371\/journal.pntd.0000727"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1718","DOI":"10.1007\/s00018-009-9053-2","article-title":"Calcium imaging of muscle cells treated with snake myotoxins reveals toxin synergism and presence of acceptors","volume":"66","author":"Pizzo","year":"2009","journal-title":"Cell. Mol. Life Sci. CMLS"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2265","DOI":"10.1016\/j.bbapap.2014.09.015","article-title":"A structure-based proposal for a comprehensive myotoxic mechanism of phospholipase A2-like proteins from viperid snake venoms","volume":"1844","author":"Fernandes","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.cbi.2015.08.002","article-title":"Phospholipase A(2) enhances the endothelial cell detachment effect of a snake venom metalloproteinase in the absence of catalysis","volume":"240","author":"Bustillo","year":"2015","journal-title":"Chem.-Biol. Interact."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"766406","DOI":"10.3389\/fphar.2021.766406","article-title":"Focused Proteomics Analysis of Habu Snake (Protobothrops flavoviridis) Venom Using Antivenom-Based Affinity Chromatography Reveals Novel Myonecrosis-Enhancing Activity of Thrombin-Like Serine Proteases","volume":"12","author":"Ogawa","year":"2021","journal-title":"Front. Pharmacol."},{"key":"ref_33","unstructured":"Farrar, J., Hotez, P.J., Junghanss, T., Kang, G., Lalloo, D., and White, N.J. (2014). 75\u2014Venomous and Poisonous Animals. Manson\u2019s Tropical Infectious Diseases (Twenty-Third Edition), W.B. Saunders."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Mac\u00eado, J.K.A., Joseph, J.K., Menon, J., Escalante, T., Rucavado, A., Guti\u00e9rrez, J.M., and Fox, J.W. (2019). Proteomic analysis of human blister fluids following envenomation by three snake species in India: Differential markers for venom mechanisms of action. Toxins, 11.","DOI":"10.3390\/toxins11050246"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2421","DOI":"10.1038\/jid.2008.118","article-title":"Skin pathology induced by snake venom metalloproteinase: Acute damage, revascularization, and re-epithelization in a mouse ear model","volume":"128","author":"Escalante","year":"2008","journal-title":"J. Investig. Dermatol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Freitas-de-Sousa, L.A., Colombini, M., Lopes-Ferreira, M., Serrano, S.M.T., and Moura-da-Silva, A.M. (2017). Insights into the mechanisms involved in strong hemorrhage and dermonecrosis induced by atroxlysin-Ia, a PI-class snake venom metalloproteinase. Toxins, 9.","DOI":"10.3390\/toxins9080239"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"5120","DOI":"10.1021\/pr900489m","article-title":"Wound exudate as a proteomic window to reveal different mechanisms of tissue damage by snake venom toxins","volume":"8","author":"Escalante","year":"2009","journal-title":"J. Proteome Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1016\/j.toxicon.2009.01.038","article-title":"Experimental pathology of local tissue damage induced by Bothrops asper snake venom","volume":"54","author":"Rucavado","year":"2009","journal-title":"Toxicon"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Pidde-Queiroz, G., Magnoli, F.C., Portaro, F.C., Serrano, S.M., Lopes, A.S., Paes Leme, A.F., van den Berg, C.W., and Tambourgi, D.V. (2013). P-I snake venom metalloproteinase is able to activate the complement system by direct cleavage of central components of the cascade. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002519"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Cruz, L.J., Luo, S., and Gopalakrishnakone, P. (2017). Inflammatory action of secretory phospholipases A2 from snake venoms. Toxins and Drug Discovery, Springer.","DOI":"10.1007\/978-94-007-6452-1"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1016\/j.bbrc.2012.11.129","article-title":"Why myotoxin-containing snake venoms possess powerful nucleotidases?","volume":"430","author":"Caccin","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Rucavado, A., Nicolau, C.A., Escalante, T., Kim, J., Herrera, C., Guti\u00e9rrez, J.M., and Fox, J.W. (2016). Viperid envenomation wound exudate contributes to increased vascular permeability via a DAMPs\/TLR-4 mediated pathway. Toxins, 8.","DOI":"10.3390\/toxins8120349"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2156","DOI":"10.1007\/s00894-014-2156-1","article-title":"Homology modeling, molecular dynamics and atomic level interaction study of snake venom 5\u2019 nucleotidase","volume":"20","author":"Arafat","year":"2014","journal-title":"J. Mol. Model."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Lazarovici, P., Marcinkiewicz, C., and Lelkes, P.I. (2019). From Snake Venom\u2019s Disintegrins and C-Type Lectins to Anti-Platelet Drugs. Toxins, 11.","DOI":"10.3390\/toxins11050303"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Suntravat, M., Sanchez, O., Reyes, A., Cirilo, A., Ocheltree, J.S., Galan, J.A., Salazar, E., Davies, P., and Sanchez, E.E. (2021). Evaluation of Signaling Pathways Profiling in Human Dermal Endothelial Cells Treated by Snake Venom Cysteine-Rich Secretory Proteins (svCRiSPs) from North American Snakes Using Reverse Phase Protein Array (RPPA). Toxins, 13.","DOI":"10.3390\/toxins13090613"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Tasoulis, T., and Isbister, G.K. (2017). A Review and Database of Snake Venom Proteomes. Toxins, 9.","DOI":"10.3390\/toxins9090290"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1146\/annurev-anchem-071015-041550","article-title":"Progress in Top-Down Proteomics and the Analysis of Proteoforms","volume":"9","author":"Toby","year":"2016","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1016\/j.jasms.2010.01.025","article-title":"Confident assignment of intact mass tags to human salivary cystatins using top-down Fourier-transform ion cyclotron resonance mass spectrometry","volume":"21","author":"Ryan","year":"2010","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1002\/jms.1242","article-title":"Snake venomics. Strategy and applications","volume":"42","author":"Calvete","year":"2007","journal-title":"J. Mass Spectrom."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/14\/9\/643\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:33:44Z","timestamp":1760142824000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/14\/9\/643"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,18]]},"references-count":49,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["toxins14090643"],"URL":"https:\/\/doi.org\/10.3390\/toxins14090643","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,18]]}}}