欢迎登录材料期刊网

材料期刊网

高级检索

基于亲和毛细管电泳,定性比较了血浆、肠道及细胞线粒体中所选的9种重要功能蛋白质与神经毒性生物毒素———软骨藻酸( domoic acid,DA)的相互作用。以 DA淌度比变化量ΔM 与蛋白质浓度 L 作图,根据斜率值大小可比较 DA与蛋白质作用的相对强弱。结果如下:可与 DA相互作用的有6种蛋白质,作用强弱为:人凝血酶>细胞色素 C≈胰蛋白酶≈免疫球蛋白 E( Ig E)≈核糖核酸酶 A>λ核酸外切酶;而铁蛋白、转铁蛋白、凝集素与 DA未表现出相互作用和亲和力。实验表明,亲和毛细管电泳具有高效、快速、所需样品量低的优点,可用于DA作用靶蛋白的筛选,为 DA的毒性机制研究和毒性防护提供基础信息。

Domoic acid( DA)is a biological neurotoxin that causes amnesic shellfish poison-ing. Study of the interactions between DA and important functional proteins contributes to understand the toxicity mechanism of DA to biological macromolecules. In this paper,the inter-actions between DA and nine important proteins in plasma,intestine and mitochondria were qualitatively compared by affinity capillary electrophoresis. Proteins were used as affinity lig-ands while DA as the affinity receptor. Proteins with the concentrations of 0,0. 2,0. 4,0. 6, 0. 8 μmol/L were added in the electrophoresis buffer and the migration times of 0. 2 mg/mL DA were detected. Then the linear graphs of the variation of DA mobility ratio(ΔM)with the pro-tein mass concentration( L)were drawn. According to the slope value,the relative strength of the interactions between DA and proteins was compared. The results showed that six proteins can interact with DA and the relative strength order was human thrombin>cytochrome C≈tryp-sin≈immunoglobulin E( Ig E)≈ribonuclease A>λ exonuclease,while ferritin,transferrin and lectin had no affinity with DA. With the advantages of high efficiency,fast analysis and less sample consumption,affinity capillary electrophoresis is a convenient method for screening DA target proteins,which will provide basic information for the toxic mechanism and defence of DA.

参考文献

[1] Quilliam M A.[J].Journal of Chromatography A,20031000(1/2):527.
[2] Linskens H F;Jackson J F.Plant Toxin Analysis[M].Berlin:Springer-Verlag,1992:185.
[3] 高利利;程金平;刘元嫄;王茜;王文华.软骨藻酸胶体金免疫层析检测试纸条的研制[J].环境科学,2011(8):2492-2496.
[4] 王茜;程金平;高利利;董宇;王文华.记忆缺失性贝毒软骨藻酸的污染现状及检测技术[J].安徽农业科学,2011(26):16070-16073,16136.
[5] 吴映璇;林峰;林海丹;邵琳智.快速高分离度液相色谱法测定贝类中软骨藻酸残留量[J].分析试验室,2008(z2):173-175.
[6] Zhao JY.;Quilliam MA.;Thibault P..ANALYSIS OF DOMOIC ACID AND ISOMERS IN SEAFOOD BY CAPILLARY ELECTROPHORESIS[J].Electrophoresis: The Official Journal of the International Electrophoresis Society,19972(2):268-276.
[7] Kvasnicka F;Sevcik R;Voldrich M.Determination of domoic acid by on-line coupled capillary isotachophoresis with capillary zone electrophoresis[J].Journal of chromatography, A: Including electrophoresis and other separation methods,20061/2(1/2):255-258.
[8] 李大志;祝文君;宋文斌;林炳承.记忆缺失性贝类毒素的主要成分--软骨藻酸的毛细管电泳分析[J].色谱,2002(2):125-128.
[9] 卫锋;王竹天.毛细管电泳法分析贝类食品中的软骨藻酸[J].中国食品卫生杂志,2003(2):107-110.
[10] 高虹;陈西平.118 软骨藻酸的毒性作用机制研究概况[J].国外医学(卫生学分册),2002(5):297-299,309.
[11] 吴冬梅;张雁秋;程伟;陆军;郑元林.软骨藻酸神经毒性作用机制研究进展[J].生态毒理学报,2013(1):1-6.
[12] Lefebvre KA;Tilton SC;Bammler TK;Beyer RP;Srinouanprachan S;Stapleton PL;Farin FM;Gallagher EP.Gene expression profiles in zebrafish brain after acute exposure to domoic acid at symptomatic and asymptomatic doses.[J].Toxicological sciences: An official journal of the Society of Toxicology,20091(1):65-77.
[13] Pérez-Gómez A;Tasker R A.[J].BMC Neuroscience,201314:72.
[14] Tsunekawa K;Kondo F;Okada T.[J].BMC Neuroscience,201314:4.
[15] Carvalho Pinto-Silva C R;Moukha S;Matias W G.[J].ENVIRONMENTAL TOXICOLOGY,200823(6):657.
[16] Liu X P;Du Y X.[J].EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY,201045(9):4043.
[17] 赵新颖;郭淑元;陈凡;屈锋;罗爱芹.毛细管电泳法表征离子液体与蛋白质的相互作用[J].分析化学,2013(8):1204-1208.
上一张 下一张
上一张 下一张
计量
  • 下载量()
  • 访问量()
文章评分
  • 您的评分:
  • 1
    0%
  • 2
    0%
  • 3
    0%
  • 4
    0%
  • 5
    0%