TePhe, a tellurium-containing phenylalanine mimic, allows monitoring of protein synthesis in vivo with mass cytometry
TePhe, a tellurium-containing phenylalanine mimic, allows monitoring of protein synthesis in vivo with mass cytometry
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DOI：
10.1073/pnas.1821151116
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发表时间：
2019-04-23
影响因子：
11.1
通讯作者：
Nitz, Mark
中科院分区：
文献类型：
--
作者：
Bassan, Jay;Willis, Lisa M.;Nitz, Mark
蛋白质合成是维持细胞内稳态的核心，其研究对于理解真核系统的功能和功能障碍至关重要。在这里，我们报告L-2-tellurienylalanine（TePhe）作为一个非典型的氨基酸，直接测量蛋白质的合成。TePhe是合成可获得的，无毒的，在生物条件下稳定的，并且碲原子允许其用质谱细胞术直接检测，而无需实验后标记。TePhe标记与苯丙氨酸竞争，但不与其他大型和芳香族氨基酸竞争，证明其作为苯丙氨酸模拟物的分子特异性;标记也在体外和体内被蛋白质合成抑制剂放线菌酮废除，验证TePhe作为翻译报告基因。在体内，用TePhe成像质谱细胞术可视化小鼠肠道、脑和肿瘤中的翻译动力学。TePhe作为蛋白质合成探针的强大性能，加上其使用的操作简单性，表明TePhe可以成为一种广泛应用的分子，用于测量体外和体内翻译。
Protein synthesis is central to maintaining cellular homeostasis and its study is critical to understanding the function and dysfunction of eukaryotic systems. Here we report L-2-tellurienylalanine (TePhe) as a noncanonical amino acid for direct measurement of protein synthesis. TePhe is synthetically accessible, nontoxic, stable under biological conditions, and the tellurium atom allows its direct detection with mass cytometry, without postexperiment labeling. TePhe labeling is competitive with phenylalanine but not other large and aromatic amino acids, demonstrating its molecular specificity as a phenylalanine mimic; labeling is also abrogated in vitro and in vivo by the protein synthesis inhibitor cycloheximide, validating TePhe as a translation reporter. In vivo, imaging mass cytometry with TePhe visualizes translation dynamics in the mouse gut, brain, and tumor. The strong performance of TePhe as a probe for protein synthesis, coupled with the operational simplicity of its use, suggests TePhe could become a broadly applied molecule for measuring translation in vitro and in vivo.