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Proteomic characterization of six Taiwanese snake venoms: Identification of species-specific proteins and development of a SISCAPA-MRM assay for cobra venom factors.
J Proteomics. 2018 09 15; 187:59-68.JP

Abstract

Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus, Daboia russelii siamensis, Bungarus multicinctus and Naja atra are the six medically important venomous snake species in Taiwan. In this study, we characterized and compared their venom protein profiles using proteomic approaches. The major snake venom proteins were identified by GeLC-MS/MS and the total venom proteome was characterized by in-solution digestion coupled with LC-MS/MS. A total of 27-52 proteins, categorized into 23 protein families, were identified in each snake's venom. The major venom components found in Viperidae species (D. acutus, T. stejnegeri, P. mucrosquamatus and D. russelii) were C-type lectin, snake venom serine proteinase, venom metalloproteinase and phospholipase A2, whereas three-finger toxin and phospholipase A2 were the major components detected in the venom of Elapidae snakes (B. multicinctus and N. atra). This study also provided the first demonstration of some low-abundance proteins in these six snake venoms, including 5'-nucleotidase, glutaminyl-peptide cyclotransferase and phosphodiesterase, among others. Furthermore, we found that cobra venom factor (CVF) is a cobra-specific protein. We produced anti-peptide antibodies against CVF and used it to develop a highly sensitive SISCAPA-MRM assay for quantifying CVF. The limit of detection and lower limit of quantification were 3.2 and 9.6 attomoles, respectively. This assay was used to precisely quantify CVF in 1 μg crude venom proteins from three Naja species and king cobra. The amount of CVF varied from 0.9 to 54.36 femtomoles (equivalent to 0.16-10.03 mg/g of venom protein).

BIOLOGICAL SIGNIFICANCE

There are six medically significant venomous snakes in Taiwan. The venoms of the four Viperidae species (Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus and Daboia russelii siamensis) cause local tissue swelling; this symptom is also seen in N. atra envenomation in humans, potentially complicating the differential diagnosis of envenomation by N. atra and Viperidae species. Thus, characterization of the venom proteomes of the six Taiwanese snakes, including the relative abundance of the major components and species-specific protein(s) in each venom type, could be useful for future venom research, including the development of new assay(s) for detecting snake species-specific venom protein(s) and new type(s) of antivenom.

Authors+Show Affiliations

Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.Department of Emergency Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.Molecular Medicine Research Center, Chang Gung University, Taoyuan 333, Taiwan; Liver Research Center, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.Department of Emergency Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan; Molecular Medicine Research Center, Chang Gung University, Taoyuan 333, Taiwan; Liver Research Center, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan; Research Center for Food and Cosmetic Safety, Research Center for Chinese Herbal Medicine, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan. Electronic address: yusong@mail.cgu.edu.tw.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29929037

Citation

Liu, Chien-Chun, et al. "Proteomic Characterization of Six Taiwanese Snake Venoms: Identification of Species-specific Proteins and Development of a SISCAPA-MRM Assay for Cobra Venom Factors." Journal of Proteomics, vol. 187, 2018, pp. 59-68.
Liu CC, Lin CC, Hsiao YC, et al. Proteomic characterization of six Taiwanese snake venoms: Identification of species-specific proteins and development of a SISCAPA-MRM assay for cobra venom factors. J Proteomics. 2018;187:59-68.
Liu, C. C., Lin, C. C., Hsiao, Y. C., Wang, P. J., & Yu, J. S. (2018). Proteomic characterization of six Taiwanese snake venoms: Identification of species-specific proteins and development of a SISCAPA-MRM assay for cobra venom factors. Journal of Proteomics, 187, 59-68. https://doi.org/10.1016/j.jprot.2018.06.003
Liu CC, et al. Proteomic Characterization of Six Taiwanese Snake Venoms: Identification of Species-specific Proteins and Development of a SISCAPA-MRM Assay for Cobra Venom Factors. J Proteomics. 2018 09 15;187:59-68. PubMed PMID: 29929037.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Proteomic characterization of six Taiwanese snake venoms: Identification of species-specific proteins and development of a SISCAPA-MRM assay for cobra venom factors. AU - Liu,Chien-Chun, AU - Lin,Chih-Chuan, AU - Hsiao,Yung-Chin, AU - Wang,Po-Jung, AU - Yu,Jau-Song, Y1 - 2018/06/19/ PY - 2018/04/10/received PY - 2018/05/28/revised PY - 2018/06/13/accepted PY - 2018/6/22/pubmed PY - 2018/6/22/medline PY - 2018/6/22/entrez KW - Cobra venom factor KW - SISCAPA-MRM KW - Taiwanese venomous snakes KW - Venom proteome SP - 59 EP - 68 JF - Journal of proteomics JO - J Proteomics VL - 187 N2 - : Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus, Daboia russelii siamensis, Bungarus multicinctus and Naja atra are the six medically important venomous snake species in Taiwan. In this study, we characterized and compared their venom protein profiles using proteomic approaches. The major snake venom proteins were identified by GeLC-MS/MS and the total venom proteome was characterized by in-solution digestion coupled with LC-MS/MS. A total of 27-52 proteins, categorized into 23 protein families, were identified in each snake's venom. The major venom components found in Viperidae species (D. acutus, T. stejnegeri, P. mucrosquamatus and D. russelii) were C-type lectin, snake venom serine proteinase, venom metalloproteinase and phospholipase A2, whereas three-finger toxin and phospholipase A2 were the major components detected in the venom of Elapidae snakes (B. multicinctus and N. atra). This study also provided the first demonstration of some low-abundance proteins in these six snake venoms, including 5'-nucleotidase, glutaminyl-peptide cyclotransferase and phosphodiesterase, among others. Furthermore, we found that cobra venom factor (CVF) is a cobra-specific protein. We produced anti-peptide antibodies against CVF and used it to develop a highly sensitive SISCAPA-MRM assay for quantifying CVF. The limit of detection and lower limit of quantification were 3.2 and 9.6 attomoles, respectively. This assay was used to precisely quantify CVF in 1 μg crude venom proteins from three Naja species and king cobra. The amount of CVF varied from 0.9 to 54.36 femtomoles (equivalent to 0.16-10.03 mg/g of venom protein). BIOLOGICAL SIGNIFICANCE: There are six medically significant venomous snakes in Taiwan. The venoms of the four Viperidae species (Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus and Daboia russelii siamensis) cause local tissue swelling; this symptom is also seen in N. atra envenomation in humans, potentially complicating the differential diagnosis of envenomation by N. atra and Viperidae species. Thus, characterization of the venom proteomes of the six Taiwanese snakes, including the relative abundance of the major components and species-specific protein(s) in each venom type, could be useful for future venom research, including the development of new assay(s) for detecting snake species-specific venom protein(s) and new type(s) of antivenom. SN - 1876-7737 UR - https://www.unboundmedicine.com/medline/citation/29929037/Proteomic_characterization_of_six_Taiwanese_snake_venoms:_Identification_of_species_specific_proteins_and_development_of_a_SISCAPA_MRM_assay_for_cobra_venom_factors_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1874-3919(18)30247-1 DB - PRIME DP - Unbound Medicine ER -