Determination of binding affinities between nuclear localization signal peptides of irradiated EGFR and KPNA using single-molecule pull-down assay and development of assay system for inhibitor screening,Bulletin of the Korean Chemical Society

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Determination of binding affinities between nuclear localization signal peptides of irradiated EGFR and KPNA using single-molecule pull-down assay and development of assay system for inhibitor screening
Bulletin of the Korean Chemical Society ( IF 2.3 ) Pub Date : 2023-05-15 , DOI: 10.1002/bkcs.12714
Jiwon Oh ₁ , Namwuk Park ₁ , Seung Wook Ham ₁

Affiliation

Epidermal growth factor receptor (EGFR) is transported into the nucleus during irradiation therapy and can cause the radiation resistance. It has been shown that irradiation activates EGFR transport to the nuclear by karyopherin alpha (KPNA) in the classical pathway, whereas ligand-activated EGFR binds with karyopherin subunit beta 1 (KPNB1). Phosphorylation of T654 located within the nuclear localization signal (NLS) of EGFR is known to activate radiation-induced nuclear EGFR transport. Therefore, we investigated the binding affinities between the NLS peptides of EGFR (⁶⁴⁵RRRHIVRKRXLRR⁶⁵⁷) and KPNA using a single-molecule pull-down assay in this study. We also showed the possible assay system for developing small molecules to modulate nuclear EGFR transport by high throughput screening, which potentially reduces the resistance during radiation therapy.

中文翻译：

使用单分子 Pull-down 测定法测定经照射的 EGFR 和 KPNA 核定位信号肽之间的结合亲和力并开发抑制剂筛选测定系统

表皮生长因子受体（EGFR）在放射治疗过程中转运至细胞核，可引起放射抵抗。研究表明，辐射激活 EGFR 通过经典途径中的核转运蛋白 α (KPNA) 转运至核，而配体激活的 EGFR 与核转运蛋白亚基 β 1 (KPNB1) 结合。已知位于 EGFR 核定位信号 (NLS) 内的 T654 磷酸化可激活辐射诱导的核 EGFR 转运。因此，我们研究了 EGFR 的 NLS 肽之间的结合亲和力 ( ⁶⁴⁵ RRRHIVRKR X LRR ⁶⁵⁷）和 KPNA 在本研究中使用单分子 Pull-down 测定。我们还展示了通过高通量筛选开发小分子来调节核 EGFR 转运的可能检测系统，这可能会降低放射治疗期间的耐药性。

更新日期：2023-05-15

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