The mass spectrometry of purified lipovitellin sample from zebrafish embryo lysate

Our preliminary research found that lipovitellin (LV) is an abundant DNA-binding protein in early zebrafish embryos. LV is the main component of vitellogenin and the largest protein of vitellogenin related products. LV is traditionally regarded as an energy reserve for developing embryos and has been shown to be involved in immune defense of developing embryos in fish. Here, purified Lv was obtained from 0 h zebrafish embryo lysate using Sephadex G-200 column combined with QAE-Sephadex A50 anion exchange column. The purified protein sample was analyzed with Easy-nLC 1200 high performance liquid chromatography (Shanghai Luming Biotechnology Co., Ltd., China). We found that although the results of mass spectrometry showed the presence of 69 proteins in the sample, LV accounted for 99.22 % of them. Table 1 shows all the protein types detected by mass spectrometry.

我们的初步研究表明，卵黄脂磷蛋白（lipovitellin，LV）是斑马鱼早期胚胎中含量丰富的DNA结合蛋白。LV是卵黄蛋白原的主要组成成分，同时也是卵黄蛋白原相关产物中分子量最大的蛋白质。传统观点认为LV为发育中的胚胎提供能量储备，已有研究表明其参与鱼类发育胚胎的免疫防御过程。本研究通过葡聚糖凝胶G-200柱联合QAE葡聚糖A50阴离子交换柱，从受精后0小时的斑马鱼胚胎裂解液中纯化得到LV。采用Easy-nLC 1200高效液相色谱仪（中国上海鹿明生物技术有限公司）对纯化后的蛋白质样品进行分析。质谱结果显示，该样品中共检测到69种蛋白质，其中LV占比达99.22%。表1列出了质谱检测到的所有蛋白质种类。