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Author:
Date:
2015-03-20
[Abstract] KRAS is the oncogene most frequently mutated in human solid tumors especially in pancreas, colon, small intestine, biliary tract and lung. We have recently demonstrated that oncogenic KRAS needs S181 phosphorylation to fully display its oncogenic features suggesting its inhibition as a therapeutic treatment against KRAS-driven tumors. Due to the importance to detect KRAS phosphorylation in human tumors and the absence of specific antibodies against phosphorylated KRAS, we developed a new protocol based on the Phos-tag SDS methodology to detect this post-translational modification for KRAS. Phos-tag is a molecule that binds specifically to phosphorylated proteins, decreasing their migration speed in SDS-PAGE and allowing its separation from the non-phosphorylated forms.
[摘要] KRAS是在人实体瘤特别是在胰腺，结肠，小肠，胆道和肺中最常突变的癌基因。 我们最近证明，致癌KRAS需要S181磷酸化充分显示其致癌特征，表明其作为对KRAS驱动的肿瘤的治疗性治疗的抑制作用。 由于在人类肿瘤中检测KRAS磷酸化的重要性和不存在针对磷酸化KRAS的特异性抗体，我们开发了基于Phos-tag SDS方法的新方案以检测这种KRAS的翻译后修饰。 Phos标签是特异性结合磷酸化蛋白质的分子，降低其在SDS-PAGE中的迁移速度并允许其从非磷酸化形式分离。
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