Anglostatin-like activity of a monoclonal antibody to the catalytic subunit of F1F0 ATP synthase
Anglostatin-like activity of a monoclonal antibody to the catalytic subunit of F1F0 ATP synthase
复制标题
DOI：
10.1158/0008-5472.can-06-1094
复制
发表时间：
2007-05-15
期刊：
影响因子：
11.2
通讯作者：
Pizzo, Salvatore V.
中科院分区：
文献类型：
--
作者：
Chi, Sulene L.;Wahl, Miriam L.;Pizzo, Salvatore V.
抗血管生成蛋白血管抑素抑制内皮细胞表面的ATP合酶，阻断细胞增殖。为了研究这种相互作用的特异性，我们产生了针对ATP合酶的单克隆抗体（mAb）。针对ATP合酶β-催化亚基的mAb（MAb 3D 5AB 1）抑制ATP合酶F-1结构域的活性并识别ATP合酶的催化β-亚基。我们将MAb 3D 5AB 1的抗体识别位点定位在含有该亚基活性位点的结构域中。MAb 3D 5AB 1也结合纯化的大肠杆菌F-1的亲和力比血管抑素对这种蛋白质的亲和力高25倍。与血管抑素类似，MAb 3D 5AB 1在酸性条件下抑制内皮细胞表面上ATP合酶产生ATP，酸性条件是细胞表面ATP合酶表现出更大活性的典型肿瘤微环境。MAb 3D 5AB 1破坏管形成，降低细胞内pH值在内皮细胞暴露于低细胞外pH值的血管抑素，也没有MAb 3D 5AB 1在角膜新生血管形成试验中显示出抗血管生成的作用，但是，两者都是有效的鸡绒毛尿囊膜试验的低pH值环境。因此，MAb 3D 5AB 1显示出上级血管抑素的血管抑素样性质，并可用于癌症化疗。
The antiangiogenic protein angiostatin inhibits ATP synthase on the endothelial cell surface, blocking cellular proliferation. To examine the specificity of this interaction, we generated monoclonal antibodies (mAb) directed against ATP synthase. mAb directed against the beta-catalytic subunit of ATP synthase (MAb3D5AB1) inhibits the activity of the F-1 domain of ATP synthase and recognizes the catalytic beta-subunit of ATP synthase. We located the antibody recognition site of MAb3D5AB1 in domains containing the active site of the subunit. MAb3D5AB1 also binds to purified Escherichia coli F-1 with an affinity 25-fold higher than the affinity of angiostatin for this protein. Mab3D5AB1 inhibits the hydrolytic activity of F-1 ATP synthase at lower concentrations than angiostatin Like angiostatin, MAb3D5AB1 inhibits ATP generation by ATP synthase on the endothelial cell surface in acidic conditions, the typical tumor microenvironment where cell surface ATP synthase exhibits greater activity. MAb3D5AB1 disrupts tube formation and decreases intracellular pH in endothelial cells exposed to low extracellular pH Neither angiostatin nor MAb3D5AB1 showed an antiangiogenic effect in the corneal neovascularization assay; however, both were effective in the low-pH environment of the chicken chorioallantoic membrane assay. Thus, MAb3D5AB1 shows angiostatin-like properties superior to angiostatin and may be exploited in cancer chemotherapy.