重组绿脓杆菌外毒素A受体结合区蛋白的初步复性及细胞生物学活性检测

Preliminary Renaturation and Detection of Cytobiological Activity of Recombinant Receptor Binding Protein of Exotoxin A of P.aeruginosa

用逐步稀释透析法将纯化的大肠杆菌表达的重组绿脓杆菌外毒素（ＰＥＡ）受体结合区蛋白初步复性，复性后的可溶性蛋白用于竞争抑制ＰＥＡ的细胞毒性实验。将１０ｎｇ／ｍｌ的ＰＥＡ与其敏感细胞Ｌ９２９共孵育，３６ｈ左右可见细胞发生病变死亡，而同时加入复性的重组ＰＥＡ受体结合区蛋白的培养细胞孔与未加ＰＥＡ阴性对照培养细胞一样仍分裂增殖，这种ＰＥＡ细胞毒性抑制作用可持续到所有培养细胞老化死亡。由此可见，重组ＰＥＡ受体结合区蛋白可在体外竞争抑制ＰＥＡ对Ｌ９２９细胞的毒性。

おhe purified recombinant receptor binding protein of exotoxin A of P.aeruginosa expressed in E.coli was renatured preliminarily by diluted progressively dialysis and the renatured solulable protein was used in the experiment of competitive inhibition of cytotoxicity of PEA.When 10ng/ml PEA were incubated with its senitive cell line L929,the cytobilogical pathological change could be found under microscope since 36h, but the cell which incubated PEA and renatured recominant receptor binding of PEA was same as the cell incubated without PEA,which was still viabile,divison and proliferation until all the cells were aged and died.The cytotoxicity of PEA could be in hibited by the recombinant receptor binding protein of PEA,the mechanism of the action is necessary to be further studied.