...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Synthesis and Evaluation of a Library of Fluorescent Dipeptidomimetic Analogues as Substrates for Modified Bacterial Ribosomes
【24h】

Synthesis and Evaluation of a Library of Fluorescent Dipeptidomimetic Analogues as Substrates for Modified Bacterial Ribosomes

机译：荧光拟肽类似物库的合成和评价作为修饰细菌核糖体的底物

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

Described herein are the synthesis and photophysical characterization of a library of aryl-substituted oxazole- and thiazole-based dipeptidomimetic analogues, and their incorporation into position 66 of green fluorescent protein (GFP) in lieu of the natural fluorophore. These fluorescent analogues resemble the fluorophore formed naturally by GFP. As anticipated, the photophysical properties of the analogues varied as a function of the substituents at the para position of the phenyl ring. The fluorescence emission wavelength maxima of compounds In the library varied from similar to 365 nm (near-UV region) to similar to 490 nm (visible region). The compounds also exhibited a large range of quantum yields (0.01-0.92). The analogues were used to activate a suppressor tRNA(CUA) and were incorporated into position 66 of GFP using an in vitro protein biosynthesizing system that employed engineered ribosomes selected for their ability to incorporate dipeptides. Four analogues with interesting photophysical properties and reasonable suppression yields were chosen, and the fluorescent proteins (FPs) containing these fluorophores were prepared on a larger scale for more detailed study. When the FPs were compared with the respective aminoacyl-tRNAs and the actual dipeptide analogues, the FPs exhibited significantly enhanced fluorescence intensities at the same concentrations. Part of this was shown to be due to the presence of the fluorophores as an intrinsic element of the protein backbone. There were also characteristic shifts in the emission maxima, indicating the environmental sensitivity of these probes. Acridon-2-ylalanine and oxazole la were incorporated into positions 39 and 66 of GFP, respectively, and were shown to form an efficient Forster resonance energy transfer (FRET) pair, demonstrating that the analogues can be used as FRET probes.
机译：本文描述了芳基取代的噁唑和噻唑基二肽模拟物库的合成和光物理表征，以及它们掺入绿色荧光蛋白（GFP）的第66位以代替天然荧光团。这些荧光类似物类似于GFP自然形成的荧光团。正如预期的那样，类似物的光物理性质随苯基环对位取代基的函数而变化。库中化合物的荧光发射波长最大值从相似的365 nm（近紫外区域）到相似的490 nm（可见光区域）不等。这些化合物还表现出较大的量子产率范围（0.01-0.92）。使用类似物激活抑制因子 tRNA （CUA），并使用体外蛋白质生物合成系统掺入 GFP 的第 66 位，该系统采用根据其掺入二肽的能力选择的工程核糖体。选取了4种具有有趣光物理性质和合理抑制产率的类似物，并更大规模地制备了含有这些荧光基团的荧光蛋白（FPs），以便进行更详细的研究。当FPs与各自的氨酰基-tRNA和实际的二肽类似物进行比较时，FPs在相同浓度下表现出显著增强的荧光强度。部分原因显示是由于荧光基团作为蛋白质骨架的内在元素的存在。发射最大值也存在特征性偏移，表明这些探头对环境的敏感性。吖啶-2-基丙氨酸和噁唑 la 分别掺入 GFP 的 39 位和 66 位，并显示出形成有效的 Forster 共振能量转移 （FRET） 对，表明类似物可用作 FRET探针。

著录项

相似文献

  • 外文文献
  • 中文文献
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