Temporal resolution of autophosphorylation for normal and oncogenic forms of EGFR and differential effects of gefitinib.
Temporal resolution of autophosphorylation for normal and oncogenic forms of EGFR and differential effects of gefitinib.
复制标题
DOI：
10.1021/bi300476v
复制
发表时间：
2012-06-26
期刊：
影响因子：
2.9
通讯作者：
Anderson KS
中科院分区：
文献类型：
--
作者：
Kim Y;Li Z;Apetri M;Luo B;Settleman JE;Anderson KS
表皮生长因子受体（EGFR）是受体酪氨酸激酶（RTK）的ErbB家族的成员。EGFR在许多不同形式的癌症中的过表达或突变突出了其作为重要治疗靶点的作用。吉非替尼是FDA批准用于治疗非小细胞肺癌（NSCLC）的第一种EGFR激酶功能小分子抑制剂，主要在EGFR中携带体细胞激酶结构域突变的肿瘤患者中显示出临床活性。在这里，我们比较了野生型EGFR自磷酸化动力学与L 834 R（也称为L 858 R）EGFR形式，这是肺癌患者中最常见的突变之一。使用快速化学淬灭、时间分辨电喷雾质谱（ESI-MS）和蛋白质印迹分析，我们检查了野生型（WT）和L 834 R EGFR中自磷酸化的顺序以及吉非替尼（Iressa ™）对单个酪氨酸磷酸化的影响。这些研究确定了参与WT EGFR下游信号传导的关键酪氨酸的自磷酸化存在时间顺序，而致癌L 834 R突变体的顺序丢失。这些研究还揭示了吉非替尼抑制酪氨酸自磷酸化的药物敏感性的独特特征模式;与EGFR的WT和致癌L 834 R突变形式不同。荧光研究表明，对于WT EGFR，吉非替尼对磷酸化蛋白的结合亲和力较弱，而对于致癌突变体L 834 R EGFR，吉非替尼的结合亲和力显著增强，可能有助于临床观察到的疗效。这种机制信息对于理解支持临床观察的分子细节以及帮助设计更有效和选择性的EGFR抑制剂非常重要。
Epidermal growth factor receptor (EGFR) is a member of the ErbB family of receptor tyrosine kinases (RTK). EGFR overexpression or mutation in many different forms of cancers has highlighted its role as an important therapeutic target. Gefitinib, the first small molecule inhibitor of EGFR kinase function to be approved for the treatment of non-small cell lung cancer (NSCLC) by the FDA, demonstrates clinical activity primarily in patients with tumors that harbor somatic kinase domain mutations in EGFR. Here, we compare wild-type EGFR autophosphorylation kinetics to the L834R (also called L858R) EGFR form, one of the most common mutations in lung cancer patients. Using rapid chemical quench, time resolved electrospray mass spectrometry (ESI-MS) and western blot analyses, we examined the order of autophosphorylation in wild-type (WT) and L834R EGFR and the effect of gefitinib (Iressa ™) on the phosphorylation of individual tyrosines. These studies establish that there is a temporal order of autophosphorylation of key tyrosines involved in downstream signaling for WT EGFR and a loss of order for the oncogenic L834R mutant. These studies also reveal unique signature patterns of drug sensitivity for inhibition of tyrosine autophosphorylation by gefitinib; distinct for WT and oncogenic L834R mutant forms of EGFR. Fluorescence studies show that for WT EGFR, the binding affinity for gefitinib is weaker for the phosphorylated protein while for the oncogenic mutant, L834R EGFR, the binding affinity of gefitinib is substantially enhanced and likely contributes to the efficacy observed clinically. This mechanistic information is important in understanding the molecular details underpinning clinical observations as well as to aid in the design of more potent and selective EGFR inhibitors.
登录
查看更多内容
影响因子：
158.5
作者：
Lynch, TJ;Bell, DW;Haber, DA
通讯作者：
Haber, DA
DOI：
10.1016/s1044-0305(03)00206-x
发表时间：
2003-09-01
影响因子：
3.2
作者：
Guo, L;Kozlosky, CJ;Johnson, RS
通讯作者：
Johnson, RS
影响因子：
11.2
作者：
Mulloy, Roseann;Ferrand, Audrey;Settleman, Jeffrey
通讯作者：
Settleman, Jeffrey
影响因子：
16
作者：
Furdui, CM;Lew, ED;Anderson, KS
通讯作者：
Anderson, KS
影响因子：
4.8
作者：
Biscardi, JS;Maa, MC;Parsons, SJ
通讯作者：
Parsons, SJ