Miyamoto Kazuhide, Saito Kazuki
Faculty of Pharmaceutical Sciences, Department of Pharmaceutical Health Care, Himeji Dokkyo University, Hyogo, Japan.
Methods Mol Biol. 2018;1867:75-87. doi: 10.1007/978-1-4939-8799-3_6.
Ubiquitination is a sequential cascade consisting of ubiquitin-activating (E1), ubiquitin-conjugating (E2), and ubiquitin-ligating (E3) enzymes. It controls numerous processes such as protein degradation, DNA repair, and signal transduction pathways. E2 enzymes are associated with a variety of diseases such as leukemia, breast cancer, lung cancer, and colorectal cancer. To date, the monitoring of E2 activity for cancer diagnosis is challenging due to its intricate cascade reaction. To surmount this hurdle, we have recently developed a novel strategy for monitoring E2 activities. Here, we describe the concise machinery of ubiquitination with artificial RING finger proteins (ARFs) functioning as E3 enzymes. This machinery enables the simplified monitoring of E2 activities. Furthermore, our system combines a signal accumulation ion-sensitive field-effect transistor biosensor with ARFs, allowing for real-time monitoring of the pathological conditions of cancer cells. The present methodology may lead to novel diagnostic techniques for cancers.
泛素化是一个由泛素激活酶（E1）、泛素结合酶（E2）和泛素连接酶（E3）组成的连续级联反应。它控制着许多过程，如蛋白质降解、DNA修复和信号转导途径。E2酶与多种疾病相关，如白血病、乳腺癌、肺癌和结直肠癌。迄今为止，由于其复杂的级联反应，监测E2活性用于癌症诊断具有挑战性。为了克服这一障碍，我们最近开发了一种监测E2活性的新策略。在这里，我们描述了以人工泛素连接酶（ARF）作为E3酶的泛素化简明机制。这种机制能够简化对E2活性的监测。此外，我们的系统将信号累积离子敏感场效应晶体管生物传感器与ARF相结合，可实时监测癌细胞的病理状况。本方法可能会带来新的癌症诊断技术。
