留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码
x 关闭 永久关闭

应中央军委要求，2022年9月起，《药学实践杂志》将更名为《药学实践与服务》，双月刊，正文96页；2023年1月起，拟出版月刊，正文64页，数据库收录情况与原《药学实践杂志》相同。欢迎作者踊跃投稿！

发形霞水母触手提取物的蛋白稳定性及其溶血活性

常银龙,  肖良,  郑杰民,  王倩倩,  张黎明

downloadPDF
文章导航 > 药学实践与服务  > 2014 >  32(6): 434-439
常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
引用本文: 常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu
CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
Citation: CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu

发形霞水母触手提取物的蛋白稳定性及其溶血活性

doi: 10.3969/j.issn.1006-0111.2014.06.009

  • 第二军医大学海医系海洋生物技术教研室, 上海 200433

基金项目: 国家自然科学基金(41176126;81370833).

Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata

  • Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai 200433, China

  • 摘要: 目的 探讨不同环境因素和处理方法对发形霞水母(Cyanea capillata)触手提取物(tentacle extract, TE)蛋白稳定性及其溶血活性的影响。 方法 结合蛋白浓度测定、溶血活性检测和SDS-PAGE分析等方法研究不同环境因素和处理方法对TE蛋白稳定性及其溶血活性的影响。 结果 TE溶血活性呈明显的剂量依赖关系,其半数溶血分数HU50=226 μg/ml;40℃水浴1 h,能去除TE中大量杂蛋白,但仍保持显著溶血活性;TE在4℃放置28 d,其溶血活性保持稳定,25℃下3 d内其溶血活性变化不明显;pH值对TE溶血活性的影响呈钟形曲线,在pH 6.0~11.0之间活性相对稳定,pH 8.0是TE保持溶血活性的最优pH条件;不同缓冲液对TE稳定性及其溶血活性影响显著,浓度大于26%的硫酸铵溶液对TE溶血蛋白具有良好的盐析作用。 结论 40℃水浴1 h预处理可显著减少TE中非活性蛋白组分,并降低样品黏性;4℃和pH 8.0是TE保持蛋白稳定性和溶血活性的最适条件;浓度大于26%的硫酸铵盐析有利于溶血蛋白组分富集。
    Abstract: Objective To investigate the influencing factors of the protein stability and hemolytic activity of tentacle extract (TE) from the jellyfish Cyanea capillata. Methods Effects of various factors and treatments on the protein stability and hemolytic activity of TE were explored by protein detection, hemolytic assay and SDS-PAGE analysis. Results TE caused a significant and dose-dependent hemolytic effect, and the HU50 of TE against 0.5% erythrocyte suspensions from SD rats was 226 μg/ml. A 40℃ water bath for 1 hour could effectively remove the contaminating proteins in TE. TE retained hemolytic activity at 4℃ for 28 days but it was unstable when kept at 25℃ over 3 days. TE was active in the range from pH 6.0 to 11.0 and the optimum pH was 8.0. Various buffer solutions had significantly different effects on the stability and hemolytic activity of TE, and a good salting-out effect was observed on the hemolytic protein of TE while the concentration of ammonium sulfate solutions was greater than 26%. Conclusion A 40℃ water bath for 1 hour could effectively remove the contaminating proteins in TE and reduce its viscosity. The optimum conditions for maintaining stability and hemolytic activity of TE were 4℃ and pH 8.0. The salting-out effect from 26% and more ammonium sulfate solutions would be conducive to the enrichment of hemolytic protein.
  • [1] Tibballs J. Australian venomous jellyfish, envenomation syndromes, toxins and therapy[J]. Toxicon, 2006, 48(7): 830-859.
    [2] Kang C, Munawir A, Cha M, et al. Cytotoxicity and hemolytic activity of jellyfish Nemopilema nomurai(Scyphozoa: Rhizostomeae)venom[J]. Comp Biochem Physiol C Toxicol Pharmacol, 2009, 150(1): 85-90.
    [3] Helmholz H, Ruhnau C, Schutt C, et al. Comparative study on the cell toxicity and enzymatic activity of two northern scyphozoan species Cyanea capillata (L) and Cyanea lamarckii (Peron & Leslieur)[J]. Toxicon, 2007, 50(1): 53-64.
    [4] Brinkman DL, Burnell JN. Biochemical and molecular characterisation of cubozoan protein toxins[J]. Toxicon, 2009, 54(8): 1162-1173.
    [5] Suput D. In vivo effects of cnidarian toxins and venoms. Toxicon, 2009, 54(8): 1190-1200.
    [6] Xiao L, He Q, Guo YF, et al. Cyanea capillata tentacle-only extract as a potential alternative of nematocyst venom: its cardiovascular toxicity and tolerance to isolation and purification procedures[J]. Toxicon, 2009, 53(1): 146-152.
    [7] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Anal Biochem, 1976, 72:248-254.
    [8] Rocha J, Peixe L, Gomes NCM, et al. Cnidarians as a source of new marine bioactive compounds-an overview of the last decade and future steps for bioprospecting[J]. Marine Drugs, 2011, 9(10): 1860-1886.
    [9] Feng J, Yu H, Xing R, et al. Partial characterization of the hemolytic activity of the nematocyst venom from the jellyfish Cyanea nozakii kishinouye[J]. Toxicol In Vitro, 2010, 24(6): 1750-1756.
    [10] Chung JJ, Ratnapala LA, Cooke IM, et al. Partial purification and characterization of a hemolysin (CAH1) from Hawaiian box jellyfish (Carybdea alata) venom[J]. Toxicon, 2001, 39(7): 981-990.
    [11] Gusmani L, Avian M, Galil B, et al. Biologically active polypeptides in the venom of the jellyfish Rhopilema nomadica[J]. Toxicon, 1997, 35(5): 637-648.
    [12] Nomura JT, Sato RL, Ahern RM, et al. A randomized paired comparison trial of cutaneous treatments for acute jellyfish(Carybdea alata) stings[J]. Am J Emerg Med, 2002, 20(7): 624-626.
    [13] Li R, Yu H, Xing R, et al. Isolation, identification and characterization of a novel antioxidant protein from the nematocyst of the jellyfish Stomolophus meleagris[J]. Int J Biol Macromol, 2012, 51(3): 274-278.
  • [1] 张艺昕, 关欣怡, 王博宁, 闻俊, 洪战英.  二氢吡啶类钙离子拮抗药物手性分析及其立体选择性药动学研究进展 . 药学实践与服务, 2024, 42(8): 319-324. doi: 10.12206/j.issn.2097-2024.202308062
    [2] 王鹏, 陈顺, 赵逸, 高守红, 王志鹏.  卡培他滨致小鼠手足综合征模型的建立及评价 . 药学实践与服务, 2024, 42(9): 385-388, 398. doi: 10.12206/j.issn.2097-2024.202308045
    [3] 陈静, 何瑞华, 翁月, 徐熠, 刘静, 黄瑾.  基于网络药理学和分子对接技术探究定清片活性成分治疗白血病的作用机制 . 药学实践与服务, 2024, 42(11): 479-486. doi: 10.12206/j.issn.2097-2024.202401073
    [4] 赖立勇, 夏天爽, 徐圣焱, 蒋益萍, 岳小强, 辛海量.  中药青蒿抗氧化活性的谱效关系研究 . 药学实践与服务, 2024, 42(5): 203-210, 216. doi: 10.12206/j.issn.2097-2024.202211012
    [5] 冯志惠, 邓仪卿, 叶冰, 安培, 张宏, 张海军.  雀梅藤石油醚提取物诱导三阴性乳腺癌细胞凋亡的实验研究 . 药学实践与服务, 2024, 42(6): 253-259. doi: 10.12206/j.issn.2097-2024.202311055
  • 加载中
WeChat 点击查看大图
计量
  • 文章访问数:  2726
  • HTML全文浏览量:  233
  • PDF下载量:  208
  • 被引次数: 0
出版历程
  • 收稿日期:  2013-11-25
  • 修回日期:  2014-07-07

发形霞水母触手提取物的蛋白稳定性及其溶血活性

doi: 10.3969/j.issn.1006-0111.2014.06.009
  • 第二军医大学海医系海洋生物技术教研室, 上海 200433
    • 基金项目:  国家自然科学基金(41176126;81370833).
  • 收稿日期: 2013-11-25
  • 修回日期: 2014-07-07

摘要: 目的 探讨不同环境因素和处理方法对发形霞水母(Cyanea capillata)触手提取物(tentacle extract, TE)蛋白稳定性及其溶血活性的影响。 方法 结合蛋白浓度测定、溶血活性检测和SDS-PAGE分析等方法研究不同环境因素和处理方法对TE蛋白稳定性及其溶血活性的影响。 结果 TE溶血活性呈明显的剂量依赖关系,其半数溶血分数HU50=226 μg/ml;40℃水浴1 h,能去除TE中大量杂蛋白,但仍保持显著溶血活性;TE在4℃放置28 d,其溶血活性保持稳定,25℃下3 d内其溶血活性变化不明显;pH值对TE溶血活性的影响呈钟形曲线,在pH 6.0~11.0之间活性相对稳定,pH 8.0是TE保持溶血活性的最优pH条件;不同缓冲液对TE稳定性及其溶血活性影响显著,浓度大于26%的硫酸铵溶液对TE溶血蛋白具有良好的盐析作用。 结论 40℃水浴1 h预处理可显著减少TE中非活性蛋白组分,并降低样品黏性;4℃和pH 8.0是TE保持蛋白稳定性和溶血活性的最适条件;浓度大于26%的硫酸铵盐析有利于溶血蛋白组分富集。

English Abstract

常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
引用本文: 常银龙, 肖良, 郑杰民, 王倩倩, 张黎明. 发形霞水母触手提取物的蛋白稳定性及其溶血活性[J]. 药学实践与服务, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu
CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
Citation: CHANG Yinlong, XIAO liang, ZHENG Jiemin, WANG Qianqian, ZHANG Liming. Protein stability and hemolytic activity of tentacle extract from the jellyfish Cyanea capillata[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(6): 434-439. doi: 10.3969/j.issn.1006-0111.2014.06.009
shu

    全文HTML

参考文献 (13)

目录

    /

    返回文章
    返回

    版权所有：《药学实践与服务》编辑委员会

    主管、主办: 中国人民解放军海军军医大学地址: 上海市杨浦区国和路325号邮政编码: 200433

    电话: （021）81871324,81871397 E-mail: [email protected]

    网站备案号 沪ICP备05003363号-1

    本系统由 北京仁和汇智信息技术有限公司 开发    百度统计