通过阴离子交换层析和凝胶过滤层析,从黄鳝(Monopterusalbus)皮肤中得到其白蛋白,命名为MAAskin。该蛋白为单链蛋白质,相对分子质量为67×10^3,经Schiff’s试剂鉴定为非糖蛋白,其N-末端氨基酸序列为GHVKW。用纯化的黄鳝血清白蛋白(MAA-serum)为免疫原免疫大白兔,制备抗MAA-serum的多克隆抗体。经ELISA检测,该抗体与MAA-skin有强烈的免疫反应,抗体滴度为1:10000。Westernblot检测,该抗体与MAA-skin也发生阳性反应,强度与MAA-seru/n相似。这说明黄鳝皮肤中分布有大量白蛋白,推测其在黄鳝皮肤的水盐代谢、渗透压维持、吸收,转运外界环境的物质等方面具有重要作用。
The anion exchange chromatography and gel filtration chromatography were performed and skin albu- min of Monopterus albus was obtained. The protein is a single chain protein with molecular weigh of 67 ~ 103, and named MAA-skin. MAA-skin was determined as a glycoprotein by Sciff~ agent. The N-terminal amino acid sequence is GHVKW. The purified MAA-serum was used to immunize rabbits to prepare polyclonal antibody. ELISA data demonstrated that the antibody titer of the MAA-skin was up to 1 : 10 000. Western blot results indi- cated that the polyclonal antibodies raised against MAA-serum strongly cross-reacted with both MAA-serum and MAA-skin with a compatible grade. The distribution of frog albumin in frog skin suggested that it plays important roles in skin physiological functions, such as water salt metabolism, osmotic pressure maintaining and substance transporting, etc.