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颜氏大疣蛛蛛毒中多肽和蛋白质的多样性分析
Determination of Peptide and Protein Diversity in the Venom of Macrothele yani
王梦如1,2,3, 张秀清1,2,3, 王颖1,2,3, 周思彤1,2,3, 杨自忠1,2,3, 李龙星3,4, 杨志斌1,2,3*
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DOI:10.11984/j.issn.1000-7083.20190035
作者单位:1. 云南省昆虫生物医药研发重点实验室, 云南大理 671000;
2. 药用特种昆虫开发国家地方联合工程研究中心, 云南大理 671000;
3. 中国西南药用昆虫及蛛形类资源开发利用协同创新中心, 云南大理 671000;
4. 大理大学分析测试中心, 云南大理 671000
中文关键字:颜氏大疣蛛;蛛毒;多肽;蛋白质;多样性
英文关键字:Macrothele yani; spider venom; peptides; proteins; diversity
中文摘要:对颜氏大疣蛛Macrothele yani蛛毒所富含的多肽与蛋白质多样性进行探索,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和超高效液相色谱-电喷雾-四极杆-飞行时间质谱技术分离和鉴定颜氏大疣蛛蛛毒中的蛋白质和多肽,并对其相对分子质量分布多样性进行分析。结果显示:粗毒中所含蛋白质的相对分子质量主要分布在35 kDa以上。在17~135 kDa分离度较佳的共有11条电泳条带,主要集中于40~120 kDa附近;在75 kDa附近弥散着高丰度的蛋白条带,75 kDa以上的蛋白质最丰富。粗毒经色谱分离后得到超过50个色谱峰,经质谱鉴定得到121个物质成分,其中,多肽类物质的相对分子质量呈双峰式分布,21%分布在500~2 000 Da,76%分布在3 000~5 000 Da,为粗毒中多肽含量最丰富的部分,且集中于35~60 min的保留时间内被洗脱。研究结果表明,颜氏大疣蛛蛛毒中含有较为丰富的多肽和蛋白类物质,这些物质的相对分子质量分布特征与已报道的其他蜘蛛既有相似性又存在具体差异。本文展示了颜氏大疣蛛蛛毒的分子多样性,为后续该毒素的物质基础研究及药用价值开发提供参考。
英文摘要:This study investigated the diversity of peptides and proteins in the venom of Macrothele yani. In this experiment, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ultra-performance liquid chromatography-electrospray-quadrupole-time of flight mass spectrometry (UPLC-ESI-Q-TOF-MS) were used to separate and identify proteins and peptides in the venom, and the distribution of molecular mass was also analyzed. The results showed that the relative molecular mass of the proteins contained in the crude venom was mainly distributed between 17 kDa and 135 kDa, and there were 11 clearly separated electrophoretic bands with molecular mass ranged from 40 kDa to 120 kDa. Additionally, high-abundance protein bands were dispersed around 75 kDa, and the most abundant bands were found over 75 kDa. Furthermore, more than 50 chromatographic peaks were separated by UPLC and 121 constituents were identified by MS analysis. 21% peptides distributed in the range of 500 Da to 2 000 Da while 76% were distributed in the range of 3 000 Da to 5 000 Da showing a bimodal distribution. As the most abundant peptides content in the crude venom of M. yani, the 76% peptides were eluted during the retention time of 35 min to 60 min. The results show that the venom of M. yani contains abundant peptides and proteins. However, this study lays a foundation for the material basic research and drug development of the toxin in the future.
2019,38(4): 408-414 收稿日期:2019-01-21
分类号:Q51;Q95
基金项目:国家地方联合工程研究中心创新能力建设项目(云发改高技[2015]395);云南省科技创新平台建设项目(2015DG030);云南省应用基础研究重点项目(2016FA012);云南省应用基础研究青年项目(2016FD074)
作者简介:王梦如(1992-),硕士研究生,从事昆虫和动物毒素方面的研究,E-mail:15947971269@163.com
*通信作者:杨志斌,E-mail:79204431@163.com
参考文献:
黄新, 王勤, 黄勤, 等. 2007. 蜘蛛毒素的细胞毒作用研究进展[J]. 广西中医药大学学报, 109(1):82-84.
张蕊. 2007. 中国异纺蛛科和狒蛛科的分类研究(蜘蛛目:原蛛下目)[D]. 保定:河北大学.
Duan ZG, Cao R, Jiang LP, et al. 2013. A combined de novo protein sequencing and cDNA library approach to the venomic analysis of Chinese spider Araneus veventricosus[J]. Journal of Proteomics, 78(1):416-427.
Escoubas P, Rash L. 2004. Tarantulas:eight-legged pharmacists and combinatorial chemists[J]. Toxicon, 43(5):555-574.
Escoubas P, Sollod B, King GF. 2006. Venom landscapes:mining the complexity of spider venoms via a combined cDNA and mass spectrometric approach[J]. Toxicon, 47(6):650-663.
Escoubas P, Quinton L, Nicholson GM. 2008. Venomics:unravelling the complexity of animal venoms with mass spectrometry[J]. Journal of Mass Spectrometry, 43(3):279-295.
Ikonomopoulou M, King G. 2013. Natural born insect killers:spider-venom peptides and their potential for managing arthropod pests[J]. Outlooks on Pest Management, 24(1):16-19.
Klint JK, Senff S, Rupasinghe DB, et al. 2012. Spider-venom peptides that target voltage-gated sodium channels:pharmacological tools and potential therapeutic leads[J]. Toxicon, 60(4):478-491.
Liao Z, Cao J, Li SM, et al. 2010. Proteomic and peptidomic analysis of the venom from Chinese tarantula Chilobrachys jingzhao[J]. Proteomics, 7(11):1892-1907.
Shi JH, Yang ZB, Zhao Y, et al. 2018. The first description on the male of Macrothele yani, and supplemental descriptions of female (Araneae:Macrothelidae)[J]. Acta Arachnologica Sinica, 27(2):103-107.
Windley MJ, Herzig V, Dziemborowicz SA, et al. 2012. Spider-venom peptides as bioinsecticides[J]. Toxins, 4(3):191-227.
Xu X, Yin CM, Griswold CE. 2002. A new species of the spider genus Macrothele from the Gaoligong Mountains, Yunnan, China (Araneae:Hexathelidae)[J]. The Pan-Pacific Entomologist, 78(2):116-119.
Yuan CH, Jin QH, Tang X, et al. 2007. Proteomic and peptidomic characterization of the venom from the Chinese bird spider, Ornithoctonus huwena Wang[J]. Journal of Proteome Research, 6(7):2792-2801.
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