蛋白质组学技术筛选与鉴定镉诱导下褐云玛瑙螺肝脏的差异蛋白质

Selection and Identification of Differential Proteins of Liver with Proteomics Techniques in Achatina Fulica Under Induction of Cadmium

比较丙酮/TCA沉淀法和直接裂解法,优化提取与分离褐云玛瑙螺(Achatina fulica,AF)肝脏全蛋白。采用丙酮/TCA沉淀法,可获得约600个蛋白质斑点。用0.5 mg/L CdCl2溶液浸泡后的去梗小白菜喂养AF,并作为镉盐诱导AF肝脏表达应激蛋白质的实验材料。采用蛋白质组学技术筛选出由镉盐诱导AF肝脏表达的14个差异蛋白质斑点,并用肽质量指纹图谱技术(peptide mass fingerprinting,PMF)和数据库比对法初步鉴定出7种差异蛋白质,其中部分为热激蛋白(heat shock protein)、甲基转移酶(Methyltransferase)、三磷酸腺苷结合盒子转运体(ABC transporter)、钼酸盐转运子亚基(molybdate transporter subunit)和磷酸甘油酸变位酶(phosphoglycerate mutase)。差异蛋白质均参与镉盐代谢,并适合作为监测土壤或食物中镉污染程度及危害性的指示蛋白质。

In comparison with direct lysis, the precipitation analysis of acetone/trichloroacetic acid was selected to optimize and separate whole proteins from the liver tissue of Achatina fulica（AF）. By this method, approximately 600 protein spots in gel were obtained. Using a cabbage exposed by 0.05 mg/L CdCl2 as AF food, the liver was selected to reveal the stress proteins in the AF exposed to acute cadmium toxicity. Fourteen of differential protein spots in AF liver were obtained by proteomics, and then seven of these spots were further identified by both peptide mass fingerprint（PMF） and database search, which indicated that these proteins in part were identified to be heat shock protein, methyhransferase, adenosine triphosphate binding cassette （ABC） transporter, molybdate transporter subunit, and phosphoglycerate mutase. The results suggest that these significant proteins can be connected with metabolism pathway of cadmium, and with biomarkers for monitoring the contamination level of the cadmium and evaluating the crisis level both food and soil.