V-ATPase Inhibitor, pH and Cell Growth Inhibition

V-ATP 酶抑制剂、pH 值和细胞生长抑制

• 批准号: 14370741
• 负责人: OHKUMA Shoji
• 金额: $ 8.13万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370741/
• 关键词: V-ATPase; Bafilomycin; Concanamycin; Affinity Probe; Neurite OutGrowth; Apoptosis; Necrosis; Cell Growth; V-ATPase; バフィロマイシン; コンカナマイシン; プロジギオシン; 細胞死; プロトン輸送; 分化; 癌

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.

即使在巴弗洛霉素抗性细胞中，巴弗洛霉素、氨或氯喹也会增加溶酶体 pH 值，而灵菌红素则会导致细胞凋亡。这些细胞对刀刀霉素敏感，表明这些化合物的结合位点彼此不同。巴弗洛霉素和刀刀霉素从质膜外作用于细胞，结果膜不可渗透的刀刀霉素也诱导神经突生长和细胞凋亡。光亲和标记表明存在除V-ATP酶16kDa蛋白脂质亚基之外的其他蛋白质。针对V-ATP酶亚基的反义寡核苷酸的结果表明Vo蛋白脂质而非V1亚基抑制细胞生长并诱导细胞死亡（坏死）。针对 V-ATP 酶亚基的抗体也观察到了类似的细胞生长抑制作用。细胞通过凋亡而死亡。这些结果不受咪唑和氨存在的影响，表明细胞pH值与反义寡核苷酸或抗体对细胞活力的影响无关。质膜上存在细胞凋亡受体。我们从嗜热栖热菌质膜上分离并测定了V-ATP酶，并成功地显示了V-ATP酶的旋转。我们在裸鼠体内研究了新的V-ATP酶抑制剂，这些物质是否能抑制肿瘤的生长。

项目成果

Keiji Tanigaki: "In bafilomycin A1-resistant cells, bafilomycin A1 raised lysosomal pH and both prodigiosins and concanamycin A inhibited growth through apoptosis"FEBS Lett.. 537. 79-84 (2003)

Keiji Tanigaki：“在巴弗洛霉素 A1 抗性细胞中，巴弗洛霉素 A1 升高溶酶体 pH，灵菌红素和刀那霉素 A 通过细胞凋亡抑制生长”FEBS Lett.. 537. 79-84 (2003)

Arai, K., Yasuda, N., Isohashi, F., Okamoto, K., Ohkuma, S.: "Inhibition of weak-base amine-induced Isis of lysosomes by cytosol."J.Biochem.. 132. 529-534 (2002)

Arai, K.、Yasuda, N.、Isohashi, F.、Okamoto, K.、Ohkuma, S.：“胞质溶胶对弱碱胺诱导的溶酶体 Isis 的抑制。”J.Biochem.. 132. 529-

大熊 勝治(編著): "細胞生物学実験法 III"廣川書店. (2003)

大隈胜晴（主编）：《细胞生物学实验方法Ⅲ》广川书店（2003）。

Keiji Tanigaki: "In bafilomycin A1-resistant cells, bafilomycin A1 raised lysosomal pH and both prodigiosins and concanamycin A inhibited growth through apoptosis"FEBS Lett.. 537(1-3). 79-84 (2003)

Keiji Tanigaki：“在巴弗洛霉素 A1 抗性细胞中，巴弗洛霉素 A1 升高溶酶体 pH，灵菌红素和刀那霉素 A 通过细胞凋亡抑制生长”FEBS Lett.. 537(1-3)。

Ken Yokoyama: "Extended longevity of C. elegans by knocking in extra copies of hsp7OF, a homologue of mot-2 (mortalin)/mthsp70/Grp75"FEBS Lett.. 516(1-3). 53-57 (2002)

Ken Yokoyama：“通过敲入 hsp7OF 的额外拷贝来延长线虫的寿命，hsp7OF 是 mot-2 (mortalin)/mthsp70/Grp75 的同源物”FEBS Lett.. 516(1-3)。