Institute for Cardiovascular Physiology, Goethe University Frankfurt, Germany.
Biomolecular Mass Spectrometry, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany; Institute of Translational Proteomics, Biochemical/Pharmacological Centre, Philipps-Universität Marburg, Germany.
Redox Biol. 2024 Nov;77:103396. doi: 10.1016/j.redox.2024.103396. Epub 2024 Oct 16.
NADPH oxidase organizer 1 (NoxO1) is a scaffold cytoplasmic subunit of the reactive oxygen species (ROS) forming Nox1 complex and involved in angiogenesis, differentiation, and atherosclerosis. We found that overexpression of NoxO1 without simultaneous overexpression of any other component of the active Nox1 complex inhibited EGF-induced wound closure and signaling, while NoxO1 KO yielded the opposite effect. Accordingly, we hypothesize NoxO1 to exert Nox1 independent functions. Using the BioID technique, we identified ErbB2 interacting protein (Erbin) as novel interaction partner of NoxO1. Colocalization of NoxO1 with EGFR, as well as with Erbin validated this finding. EGF treatment interrupted colocalization of NoxO1 and EGFR. EGF mediated kinase activation was delayed in NoxO1 overexpressing cells, while knockout of NoxO1 had the opposite effect. In conclusion, Erbin was identified as a novel NoxO1 interacting protein. Through the subsequent interaction of NoxO1 and EGFR, NoxO1 interferes with EGF signaling. The results of this study suggest a potential role of NoxO1 as an adaptor protein with functions beyond the well-established enabling of Nox1 mediated ROS formation.
NADPH 氧化酶调节因子 1（NoxO1）是活性氧（ROS）形成 Nox1 复合物的细胞质支架亚基，参与血管生成、分化和动脉粥样硬化。我们发现，NoxO1 的过表达而没有同时过表达 Nox1 复合物的任何其他活性成分，会抑制 EGF 诱导的伤口闭合和信号转导，而 NoxO1 的敲除则产生相反的效果。因此，我们假设 NoxO1 发挥了非 Nox1 依赖性的功能。使用 BioID 技术，我们鉴定出 ErbB2 相互作用蛋白（Erbin）是 NoxO1 的新的相互作用伙伴。NoxO1 与 EGFR 以及 Erbin 的共定位验证了这一发现。EGF 处理中断了 NoxO1 和 EGFR 的共定位。在 NoxO1 过表达细胞中，EGF 介导的激酶激活被延迟，而 NoxO1 的敲除则产生相反的效果。总之，Erbin 被鉴定为 NoxO1 的一种新的相互作用蛋白。通过 NoxO1 和 EGFR 的后续相互作用，NoxO1 干扰了 EGF 信号转导。这项研究的结果表明，NoxO1 作为一种衔接蛋白，具有超越其公认的 Nox1 介导 ROS 形成功能的潜在作用。
