Citation: WANG Xiaoqian, GAO Tie, HONG Zhuan, QU Feng. Affinity capillary electrophoresis for screening proteins interacting with domoic acid[J]. Chinese Journal of Chromatography, ;2015, 33(7): 673-677. doi: 10.3724/SP.J.1123.2015.03026 shu

Affinity capillary electrophoresis for screening proteins interacting with domoic acid

WANG Xiaoqian ¹ ,
GAO Tie ¹ ,
HONG Zhuan ² ,
QU Feng ^1,,

1.
1. School of Life Science, Beijing Institute of Technology, Beijing 100081, China;

Corresponding author: QU Feng,
Received Date: 19 March 2015

Fund Project: 国家自然科学基金项目(21175011,21375008) (21175011,21375008)国家"973"计划项目(2012CB910603) (2012CB910603)国家海洋局海洋生物资源综合利用工程技术研究中心开放基金(MBRCU201304). (MBRCU201304)

Domoic acid (DA) is a biological neurotoxin that causes amnesic shellfish poisoning. 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 interactions between DA and nine important proteins in plasma, intestine and mitochondria were qualitatively compared by affinity capillary electrophoresis. Proteins were used as affinity ligands 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 protein 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≈trypsin≈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.
- affinity capillary electrophoresis,
- domoic acid,
- protein,
- interaction
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