Department of Chemistry, University of Toronto, Toronto, ON, Canada M5S 3H6.
The Campbell Family Institute for Cancer Research, Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada M5T 2M9.
Proc Natl Acad Sci U S A. 2019 Apr 23;116(17):8155-8160. doi: 10.1073/pnas.1821151116. Epub 2019 Apr 10.
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.
蛋白质合成是维持细胞内稳态的核心,对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-碲代苯丙氨酸(TePhe)作为一种非典型氨基酸,可直接测量蛋白质合成。TePhe 可通过合成获得,无毒,在生物条件下稳定,碲原子允许其通过质谱流式细胞术直接检测,无需实验后标记。TePhe 标记与苯丙氨酸竞争,但不与其他大和芳香族氨基酸竞争,证明其作为苯丙氨酸类似物的分子特异性;该标记也被体外和体内的蛋白质合成抑制剂环己酰亚胺所阻断,验证了 TePhe 作为翻译报告物的作用。在体内,使用 TePhe 的成像质谱流式细胞术可以在小鼠肠道、大脑和肿瘤中可视化翻译动力学。TePhe 作为蛋白质合成探针的出色性能,加上其使用的操作简单性,表明 TePhe 可能成为一种广泛应用于体外和体内测量翻译的分子。