问号钩端螺旋体赖株溶血素基因LA0202的溶血机制

The functional classification of a hemolysin protein LA0202 in Leptospira interrogans strain Lai

目的对问号钩端螺旋体赖株溶血素基因LA0202的溶血机制进行研究。方法纯化重组蛋白LA0202并制备多克隆抗体;Western blot验证重组蛋白在问号钩端螺旋体培养过程中的表达;并进行重组蛋白LA0202溶血活性影响因素实验。结果Western blot显示LA0202在钩体培养过程中可以表达,加入二价金属阳离子对重组蛋白溶血活性亦无明显激活作用,加入蛋白酶抑制剂对溶血活性无明显抑制作用,高温条件致使溶血活性丧失,加入渗透压保护剂PEG6000能抑制溶血活性。结论问号钩端螺旋体赖株溶血素基因LA0202为在钩体生长过程中表达的一成孔蛋白类溶血素。

Objective To investigate the hemolytic mechanism of LA0202 protein. Methods The recombinant protein LA0202 was purified and its polyclone antibodies were produced. The expression of LA0202 protein in L. interrogans strains Lai was confirmed by Western blot. Moreover, the factors which could affect the hemolytic activity of LA0202 protein were investigated. Results LA0202 protein was actively expressed in L. interrogans strains Lai. Neither divalent cations nor phospholipase inhibitors altered the hemolytic activity of LA0202 protein. Heat completely inactivated LA0202 protein. Likewise, osmotic protectants could inhibit the hemolytic activity of LA0202 protein. Conclusions The LA0202 protein actively expressed in L. interrogans strains Lai belongs to the poreforming hemolysin.