Yahiro K, Niidome T, Hatakeyama T, Aoyagi H, Kurazono H, Padilla P I, Wada A, Hirayama T
Faculty of Engineering, Nagasaki University, Nagasaki, 852, Japan.
Biochem Biophys Res Commun. 1997 Sep 18;238(2):629-32. doi: 10.1006/bbrc.1997.7345.
To investigatie a potential mechanism of how Helicobacter pylori establishes infection, we purified a lot of vacuolating toxin (VacA) from supernatant of H. pylori ATCC49503 (tox+ strain 60190). We used an antibody which was prepared by immunizing rabbits with a synthetic peptide consisting of 16 amino acids reflecting a portion (Glu69-Arg83) of amino acid sequence of Vac A. VacA caused vacuoles in human gastric cancer cell lines AZ-521 AGS, and monkey kidney cell line COS-7, but not human promyeloblastic cell line HL-60. By immunoprecipitation analysis using anti VacA antibody, a biotinylated cell surface protein of 140kDa (p140) was precipitated only when the lysates of VacA-susceptible cells were incubated with VacA but not with inactivated VacA, indicating the association of p140 with VacA.
为了研究幽门螺杆菌建立感染的潜在机制，我们从幽门螺杆菌ATCC49503（毒素阳性菌株60190）的上清液中纯化了大量空泡毒素（VacA）。我们使用了一种抗体，该抗体是通过用一种由16个氨基酸组成的合成肽免疫兔子制备的，该合成肽反映了VacA氨基酸序列的一部分（Glu69 - Arg83）。VacA在人胃癌细胞系AZ - 521、AGS以及猴肾细胞系COS - 7中引起空泡，但在人早幼粒细胞系HL - 60中未引起空泡。通过使用抗VacA抗体的免疫沉淀分析，仅当VacA敏感细胞的裂解物与VacA而非灭活的VacA孵育时，一种140kDa的生物素化细胞表面蛋白（p140）才会沉淀，这表明p140与VacA有关联。
