...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Characterization of AT(4) receptor from bovine aortic endothelium with photosensitive analogues of angiotensin IV
【24h】

Characterization of AT(4) receptor from bovine aortic endothelium with photosensitive analogues of angiotensin IV

机译：用血管紧张素IV的光敏类似物表征牛主动脉内皮中的AT（4）受体

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Newly developed photosensitive analogues of AngIV were used to characterize the AT(4) receptor of bovine aortic endothelial cells, The photoactivatable AngIV analogues [N-3-Phe(6)]AngIV and [Bpa(6)]AngIV displayed high affinities for AT(4) receptor, with IC50's of 3.7 +/- 0.3 and 19.1 +/- 3.5 nM, respectively, The radioiodinated ligands showed a good efficiency of photoaffinity labeling demonstrated by high proportions (60-75%) of acid-resistant binding. Covalently labeled receptor was solubilized under reducing or nonreducing conditions and subjected to SDS-PAGE. Under nonreducing conditions, autoradiographies revealed a major band of M-r 186 +/- 2 kDa and a minor band of M-r 241 +/- 6 kDa. The labeling of these bands was completely abolished in the presence of 10 mu M AngIV, Under reducing conditions, only the low M-r 186 kDa band was revealed. After endoglycosidase digestion with an enzyme that cleaves N-linked saccharides, the M-r of the denatured AT(4) receptor was decreased by 31% to a value of 129 +/- 10 kDa. Kinetic studies revealed a stepwise process of AT(4) receptor deglycosylation by endoglycosidase F, suggesting at least two different sites of N-linked saccharides. Mild trypsin treatment of photolabeled endothelial cell membranes released a large fragment of M-r 177 +/- 3 kDa which accounts for about 95% of the whole receptor molecular mass. These results demonstrate that [N-3-Phe(6)]AngIV and [Bpa(6)]AngIV are very efficient tools for selective photoaffinity labeling of AT(4) receptor, We have shown that AT(4) receptor is a 186 kDa integral membrane glycoprotein with a very large extracellular domain. These properties are consistent with those of a growth factor or cytokine receptor. [References: 32]
机译：AngIV的最新开发的光敏类似物用于表征牛主动脉内皮细胞的AT（4）受体。可光活化的AngIV类似物[N-3-Phe（6）] AngIV和[Bpa（6）] AngIV显示出对AT的高亲和力（4）受体，IC50分别为3.7 +/- 0.3和19.1 +/- 3.5 nM，放射性碘标记的配体表现出良好的光亲和性标记效率，高比例（60-75％）的耐酸结合性证明了这一点。将共价标记的受体在还原或非还原条件下溶解并进行SDS-PAGE。在非还原条件下，放射自显影显示M-r 186 +/- 2 kDa的主带和M-r 241 +/- 6 kDa的小带。在10μMAngIV存在下，这些条带的标记被完全取消。在还原条件下，仅显示出低的M-r 186 kDa条带。用切割N-连接糖的酶消化糖苷内切酶后，变性AT（4）受体的M-r降低31％，达到129 +/- 10 kDa。动力学研究表明，内切糖苷酶F使AT（4）受体去糖基化的逐步过程，提示了N-连接糖的至少两个不同位点。轻度胰蛋白酶处理光标记的内皮细胞膜会释放出M-r 177 +/- 3 kDa的大片段，约占整个受体分子量的95％。这些结果表明[N-3-Phe（6）] AngIV和[Bpa（6）] AngIV是用于AT（4）受体选择性光亲和标记的非常有效的工具，我们已经表明AT（4）受体是186具有非常大的细胞外结构域的kDa整合膜糖蛋白。这些性质与生长因子或细胞因子受体的性质一致。 [参考：32]

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号