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由于分泌蛋白在重大生理和病理过程中的重要角色和功能,使得分泌蛋白质组的研究备受人们关注.高效提取分泌蛋白质是分泌蛋白质组学研究的第一步,也是关键的一步.本研究首次比较了3种常用的提取方法:超滤法、沉淀法和透析法.以具有高自发性转移潜能的人肝癌细胞系LM3为研究对象,采用3种方法平行提取分泌蛋白,发现超滤法的提取效率最高;蛋白质经过溶液酶解,在线的一维反相高效液相色谱分离和ESI-MS/MS质谱鉴定,共鉴定了360个非冗余蛋白,其中有34,110和29个蛋白分别只在超滤法、沉淀法和透析法提取的样品中被鉴定到,沉淀法占优势,3种方法互相补充.360个蛋白中,有42个分泌蛋白为首次被鉴定到,扩大了已有的人肝癌转移细胞分泌蛋白数据库.
Due to the important role and function of secreted proteins in major physiological and pathological processes, the research on secretome proteomics has attracted much attention.Efficient extraction of secreted proteins is the first and crucial step in the study of secretory proteomics.In this study, Three commonly used extraction methods were compared: ultrafiltration, precipitation and dialysis.According to LM3, a human hepatocellular carcinoma cell line with high spontaneous metastatic potential, three kinds of methods were used to extract secretory proteins in parallel, And the highest extraction efficiency. The proteins were identified by solution digestion, on-line one-dimensional reversed-phase high performance liquid chromatography and ESI-MS / MS. A total of 360 non-redundant proteins were identified. Among them, 34, 110 and 29 proteins were identified Ultrafiltration, precipitation and dialysis were extracted samples were identified, the precipitation method is dominant, three kinds of methods complement each other.Of the 360 proteins, 42 secreted proteins were identified for the first time, expanding the existing people Liver cancer metastasis cell secretory protein database.