Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.
Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts.
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DOI：
10.1083/jcb.111.3.1305
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发表时间：
1990-09
期刊：
影响因子：
--
通讯作者：
Lakkakorpi P
中科院分区：
文献类型：
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作者：
Väänänen HK;Karhukorpi EK;Sundquist K;Wallmark B;Roininen I;Hentunen T;Tuukkanen J;Lakkakorpi P
从鸡髓质骨或分离的破骨细胞制备的微粒体膜囊泡被证明具有atp依赖的H(+)-运输活性。该活性对n -乙基马来酰亚胺敏感，但对寡霉素和正钒酸盐耐药，提示为液泡型atp酶。此外，用免疫印迹法分析60- kd和70-kD破骨细胞膜抗原与粗神经孢子虫空泡atp酶的免疫交叉反应性。免疫组化结果显示，鸡骨和大鼠骨的破骨细胞有相同的抗体标记。免疫电镜发现这些抗原存在于大鼠破骨细胞的尖膜和外髓质内条的肾插层细胞中。甲状旁腺激素预处理动物可增强破骨细胞顶膜的免疫反应。当使用抗胃H+、K(+)- atp酶抗体时，破骨细胞无免疫反应。这些结果表明破骨细胞通过空泡H(+)- atp酶分泌质子来吸收骨。
Microsomal membrane vesicles prepared either from chicken medullary bone or isolated osteoclasts were shown to have ATP-dependent H(+)- transport activity. This activity was N-ethylmaleimide-sensitive but resistant to oligomycin and orthovanadate, suggesting a vacuolar-type ATPase. Furthermore, immunological cross-reactivity of 60- and 70-kD osteoclast membrane antigens with Neurospora crassa vacuolar ATPase was observed when analyzed by immunoblotting. Same antibodies labeled only osteoclasts in chicken and rat bone in immunohistochemistry. Immunoelectronmicroscopy localized these antigens in apical membranes of rat osteoclasts and kidney intercalated cells of inner stripe of outer medulla. Pretreatment of animals with parathyroid hormone enhanced the immunoreaction in the apical membranes of osteoclasts. No immunoreaction was seen in osteoclasts when antibodies against gastric H+,K(+)-ATPase were used. These results suggest that osteoclast resorbs bone by secreting protons through vacuolar H(+)-ATPase.