V-ATPase Inhibitor, pH and Cell Growth Inhibition

甚至在对巴菲罗星耐药的细胞中，巴菲霉素、氨水或氯喹也能提高溶酶体的pH，而灵芝菌素可诱导细胞凋亡。这些细胞对刀豆素敏感，提示这些成分的结合部位(S)不同。Bafilmycins和Concanamyins作用于质膜外的细胞，结果表明膜不通透性Concamycins还可诱导轴突生长和细胞凋亡。光亲和标记结果表明，V-ATPase亚基中存在除16 kDa蛋白脂亚单位外的其他蛋白质(S)。与V-ATPase亚基的反义寡核苷酸杂交结果表明，Vo蛋白脂类而不是V1亚基抑制细胞生长并诱导细胞死亡(坏死)。抗V-ATPase亚基抗体对细胞生长的抑制作用类似。这些细胞通过细胞凋亡而死亡。这些结果不受咪唑和氨的存在的影响，这表明细胞的pH与反义寡核苷酸或抗体对细胞活力的影响无关。质膜上存在凋亡受体(S).我们从嗜热嗜热杆菌质膜上分离并测定了V-ATPase，并成功地显示了V-ATPase的旋转.我们还探讨了新的V-ATPase抑制剂在裸鼠体内是否能抑制肿瘤的生长.

Even in bafilomycin-resistant cells, lysosomal pH was increased by bafilomycins, ammonia or chloroquine, and prodigiosins indeced apoptosis. These cells were sensitive to concanamycins, suggesting the binding site(s) these componds are different from each other. Bafilomycins adn concanamacyns act on the cells from outside plasma membrane from theresults that membrane-impermeable concaamycins also induced neurite-outgrowth and apoptosis. Photoaffinity labelling suggested the presence of other protein(s) than 16kDa proteolipid subunit of V-ATPase.Results with antisence-oligonucleotide against V-ATPase subunits suggested that Vo proteolipids but not V1 subunit inhibited the cell growth and induced cell death(necrosis). Similar inhibitin of cell growth was observed by antibodies against V-ATPase subunits. The cells are dead through apoptosis. These results are not affected by te presence of imidazole and ammonia, suggesting that cellular pH are not responsible to the effect of antisence oligonucleotide or antibody on cellular viability. The presence of receptor(s) for apoptosis on teh plasme membrenes.We have isolated and determined the V-ATPase from the plasma membrane of Thermus thermophilis and suceeded in showing the rotation of V-ATPase.We have looked into the new inhibitors against V-ATPase in nude mice, if these substances inhibited the growth of tumors.