Sappakhaw Khomkrit, Jantarug Krittapas, Slavoff Sarah A, Israsena Nipan, Uttamapinant Chayasith
School of Biomolecular Science and Engineering Vidyasirimedhi Institute of Science and Technology (VISTEC) Rayong 21210 Thailand.
Department of Chemistry Yale University New Haven CT 06520 USA.
Angew Chem Weinheim Bergstr Ger. 2021 Feb 19;133(8):3980-3985. doi: 10.1002/ange.202010703. Epub 2020 Dec 15.
Polypeptides generated from proteolytic processing of protein precursors, or proteolytic proteoforms, play an important role in diverse biological functions and diseases. However, their often-small size and intricate post-translational biogenesis preclude the use of simple genetic tagging in their cellular studies. Herein, we develop a labeling strategy for this class of proteoforms, based on residue-specific genetic code expansion labeling with a molecular beacon design. We demonstrate the utility of such a design by creating a molecular beacon reporter to detect amyloid-β peptides, known to be involved in the pathogenesis of Alzheimer's disease, as they are produced from amyloid precursor protein (APP) along the endocytic pathway of living cells.
由蛋白质前体的蛋白水解加工产生的多肽，即蛋白水解蛋白变体，在多种生物学功能和疾病中发挥着重要作用。然而，它们通常尺寸较小且翻译后生物合成复杂，这使得在细胞研究中无法使用简单的基因标记。在此，我们基于分子信标设计的残基特异性遗传密码扩展标记，开发了一种针对这类蛋白变体的标记策略。我们通过创建一个分子信标报告基因来检测淀粉样β肽，证明了这种设计的实用性。淀粉样β肽已知参与阿尔茨海默病的发病机制，当它在活细胞的内吞途径中由淀粉样前体蛋白（APP）产生时即可被检测到。
