V-ATPase Inhibitor, pH and Cell Growth Inhibition

即使在巴非霉素耐药细胞中，巴非霉素、氨或氯喹也可使溶酶体pH升高，灵菌菌素可诱导细胞凋亡。这些细胞对康卡霉素敏感，表明这些化合物的结合位点彼此不同。巴菲尔霉素和康卡霉素作用于细胞外质膜，结果表明膜不可渗透的康卡霉素也诱导神经突生长和凋亡。光亲和标记表明，V-ATP酶中除了16 kDa蛋白脂亚基外，还存在其它蛋白质，V-ATP酶亚基的反义寡核苷酸结果表明，Vo蛋白脂而非V1亚基抑制细胞生长，并诱导细胞死亡（坏死）。V-ATP酶亚基的抗体也能抑制细胞生长。细胞通过凋亡而死亡。这些结果不受咪唑和氨的存在的影响，表明细胞的pH不是反义寡核苷酸或抗体对细胞活力的影响的原因。细胞质膜上凋亡受体的存在.我们从嗜热栖热菌和蔗糖栖热菌的质膜上分离并测定了V-ATP酶，显示了V-ATP酶的旋转.我们还在裸鼠体内研究了V-ATP酶的新抑制剂是否抑制肿瘤的生长.

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.