Araki Kazutaka, Ushioda Ryo, Kusano Hidewo, Tanaka Riko, Hatta Tomohisa, Fukui Kazuhiko, Nagata Kazuhiro, Natsume Tohru
Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Tokyo 135-0064, Japan.
Faculty of Life Sciences, Kyoto Sangyo University, Kita-Ku, Kyoto 603-8555, Japan.
Anal Biochem. 2017 Mar 1;520:22-26. doi: 10.1016/j.ab.2016.12.025. Epub 2016 Dec 31.
Thiol-based redox control is among the most important mechanisms for maintaining cellular redox homeostasis, with essential participation of cysteine thiols of oxidoreductases. To explore cellular redox regulatory networks, direct interactions among active cysteine thiols of oxidoreductases and their targets must be clarified. We applied a recently described thiol-ene crosslinking-based strategy, named divinyl sulfone (DVSF) method, enabling identification of new potential redox relay partners of the cytosolic oxidoreductases thioredoxin (TXN) and thioredoxin domain containing 17 (TXNDC17). Applying multiple methods, including classical substrate-trapping techniques, will increase understanding of redox regulatory mechanisms in cells.
基于硫醇的氧化还原控制是维持细胞氧化还原稳态的最重要机制之一,氧化还原酶的半胱氨酸硫醇起着至关重要的作用。为了探索细胞氧化还原调节网络,必须阐明氧化还原酶的活性半胱氨酸硫醇与其靶标之间的直接相互作用。我们应用了一种最近描述的基于硫醇-烯交联的策略,即二乙烯基砜(DVSF)方法,该方法能够识别胞质氧化还原酶硫氧还蛋白(TXN)和含硫氧还蛋白结构域17(TXNDC17)的新的潜在氧化还原中继伙伴。应用多种方法,包括经典的底物捕获技术,将增进对细胞中氧化还原调节机制的理解。