Department of Biochemistry, Kochi University Medical School, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
Sensors (Basel). 2012 Nov 22;12(12):16037-45. doi: 10.3390/s121216037.
Important biological events associated with plasma membranes, such as signal transduction, cell adhesion, and protein trafficking, are mediated through the membrane microdomains. We have developed a novel method termed enzyme-mediated activation of radical sources (EMARS) to identify coclustering molecules on the cell surface under living conditions, which features a radical formation from an aryl azide reagent by horseradish peroxidase (HRP). For identification of molecules labeled by the EMARS reaction, antibody array system and mass spectrometry-based proteomics approaches are available. Spatio- temporally-regulated interaction between b1 integrin and ErbB4 involved in fibronectin-dependent cell migration and therapeutic antibody-stimulated interaction between FGFR3 and CD20 were discovered using the EMARS method.
与质膜相关的重要生物学事件，如信号转导、细胞黏附和蛋白质运输，都是通过膜微区来介导的。我们开发了一种新的方法，称为酶介导的自由基源激活（EMARS），以在活细胞条件下鉴定细胞表面上的共聚类分子，其特征是通过辣根过氧化物酶（HRP）从芳基叠氮试剂中形成自由基。为了鉴定 EMARS 反应标记的分子，可以使用抗体阵列系统和基于质谱的蛋白质组学方法。使用 EMARS 方法发现了 b1 整合素和 ErbB4 之间的时空调节相互作用，该相互作用涉及纤维连接蛋白依赖性细胞迁移和治疗性抗体刺激 FGFR3 和 CD20 之间的相互作用。
