聚腺苷二磷酸核糖的基因编码荧光传感器。
Genetically Encoded Fluorescent Sensor for Poly-ADP-Ribose.
机构信息
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia.
Institute of Molecular Genetics, Kurchatova Sq. 2, 123182 Moscow, Russia.
出版信息
Int J Mol Sci. 2020 Jul 15;21(14):5004. doi: 10.3390/ijms21145004.
Poly-(ADP-ribosyl)-ation (PARylation) is a reversible post-translational modification of proteins and DNA that plays an important role in various cellular processes such as DNA damage response, replication, transcription, and cell death. Here we designed a fully genetically encoded fluorescent sensor for poly-(ADP-ribose) (PAR) based on Förster resonance energy transfer (FRET). The WWE domain, which recognizes iso-ADP-ribose internal PAR-specific structural unit, was used as a PAR-targeting module. The sensor consisted of cyan Turquoise2 and yellow Venus fluorescent proteins, each in fusion with the WWE domain of RNF146 E3 ubiquitin ligase protein. This bipartite sensor named sPARroW (sensor for PAR relying on WWE) enabled monitoring of PAR accumulation and depletion in live mammalian cells in response to different stimuli, namely hydrogen peroxide treatment, UV irradiation and hyperthermia.
聚（ADP-核糖）化（PARylation）是一种蛋白质和 DNA 的可逆翻译后修饰，在 DNA 损伤反应、复制、转录和细胞死亡等各种细胞过程中发挥着重要作用。在这里，我们设计了一种基于荧光共振能量转移（FRET）的全基因编码的聚（ADP-核糖）（PAR）荧光传感器。WWE 结构域，识别 iso-ADP-核糖内部 PAR 特定的结构单元，被用作 PAR 靶向模块。该传感器由青色 Turquoise2 和黄色 Venus 荧光蛋白组成，每个蛋白与 RNF146 E3 泛素连接酶蛋白的 WWE 结构域融合。这种二聚体传感器命名为 sPARroW（基于 WWE 的 PAR 传感器），可在活哺乳动物细胞中监测 PAR 的积累和消耗，以响应不同的刺激，即过氧化氢处理、紫外线照射和高温处理。
Zotero 插件