Studies on physiological functions of vacuolar type H^+-ATPase using low molecular weight probes

低分子探针研究液泡型H^-ATP酶的生理功能

• 批准号: 09460047
• 负责人: NAGAI Kazuo
• 金额: $ 7.3万
• 依托单位: TOKYO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460047/
• 关键词: Biologically active compound; Immunomodulator; Bone metabolism; Osteoclast; Cytotoxic Tcell; Vacuolar type H^+-ATPase; Bioprobe

Effects of concanamycin B, a specific inhibitor of vacuolar type H^+-ATPase (V-ATPase), and prodigiosin 25-C, which inhibits proton pump activity of V-ATPase without affecting its ATPase activity, on acidification of acidic intracellular granules in osteoclasts and on osteoclastic bone resorption were determined. These two inhibitors suppressed the acidification of the granules as well as bone resorption. Destruxins A, B and B, which are known to inhibit proton pump activity of V-ATPase, did not inhibit acidification of acidic intracellular granules in osteoclasts. However, they inhibited osteoclastic bone resorption by inducing morphological changes of osteoclasts that are not observed in the osteoclasts treated with concanamycin B or prodigiosin 25-C.It is clearly demonstrated that the morphological changes caused by destruxins are attributable to disruption of actin rings in osteoclasts which are essential cytoskeletal structures for bone resorption. The disruptive potency of destruxin E, which contains epoxide moiety in the molecule, was 30 times more evident than that of destruxin A and B, suggesting that the epoxide plays an important role in the expression of biological activities.

测定了空泡型H^+-ATP酶（V-ATP酶）的特异性抑制剂康卡霉素B和抑制V-ATP酶质子泵活性而不影响其ATP酶活性的灵菌红素25-C对破骨细胞酸性细胞内颗粒酸化和破骨细胞骨吸收的影响。这两种抑制剂抑制颗粒的酸化以及骨吸收。已知能抑制V-ATP酶质子泵活性的破坏素A、B和B不能抑制破骨细胞内酸性颗粒的酸化。然而，它们通过诱导破骨细胞的形态变化来抑制骨吸收，而这在用康卡霉素B或灵菌红素25-C处理的破骨细胞中未观察到。这清楚地表明，由destruxins引起的形态变化可归因于破骨细胞中肌动蛋白环的破坏，破骨细胞是骨吸收所必需的细胞骨架结构。在分子中含有环氧化物部分的破坏力比破坏A和破坏B的破坏力大30倍，表明环氧化物在生物活性的表达中起重要作用。

项目成果

Ken-ichi Togashi: "Characterization of a series of vacuolar type H^+-ATPase inhibitors on CTL-mediated cytotoxicitty" Immunol.Let.55. 139-144 (1997)

Ken-ichi Togashi：“一系列液泡型H + -ATP酶抑制剂对CTL介导的细胞毒性的表征”Immunol.Let.55。

Shin-ichi Hamada, Takao Kataoka, Je-Tae Woo, Atsushi Yamada, Takashi Yoshida, Toshio Nishimura, Noboru Otake, and Kazuo Nagai: "Immunosuppressive effects of gallic acid and chebulagic acid on CTL-mediated cytotoxicity" Biol.Pharma.Bull.20. 1017-1019 (1997

Shin-ichi Hamada、Takao Kataoka、Je-Tae Woo、Atsushi Yamada、Takashi Yoshida、Toshio Nishimura、Noboru Otake 和 Kazuo Nagai：“没食子酸和 chebulagic 酸对 CTL 介导的细胞毒性的免疫抑制作用”Biol.Pharma.Bull。

Je-Tae Woo: "Prodigiosin 25-C and metacycloprodigiosin suppress the bone resorption by osteoclasts" Biosci.Biotech.Biochem.61. 400-402 (1997)

Je-Tae Woo：“灵菌红 25-C 和变环灵菌红抑制破骨细胞的骨吸收”Biosci.Biotech.Biochem.61。

Takao Kataoka et al.: "Inactivation and proteolytic degradation of perforin wihin lytic granules upon neutralization of acidic pH" Immunology. 91. 493-500 (1997)

Takao Kataoka 等人：“中和酸性 pH 后裂解颗粒内穿孔素的失活和蛋白水解降解”免疫学。

Je-Tea Woo: "Low molecular weight microbial metabolites that suppress bone resorption by inhibiting vacuolar type proton pump activity of osteoclasts" in K.Funatsu et al. (eds.) Animal Cell Technology : Basic & Applied Aspects. 8. 627-632 (1997)

Je-Tea Woo：“低分子量微生物代谢物通过抑制破骨细胞的液泡型质子泵活性来抑制骨吸收”，K.Funatsu 等人。