Analysis of Phosphotyrosine Signaling Networks in Lung Cancer Cell Lines.
Analysis of Phosphotyrosine Signaling Networks in Lung Cancer Cell Lines.
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DOI：
10.1007/978-1-4939-7154-1_16
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发表时间：
2017
影响因子：
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通讯作者：
Malgorzata Broncel;Paul H. Huang
中科院分区：
文献类型：
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作者：
Malgorzata Broncel;Paul H. Huang
稳健分离和鉴定酪氨酸残基磷酸化的肽是破译由蛋白酪氨酸激酶（包括参与肿瘤发生的激酶）控制的复杂信号网络的关键步骤。含磷酸酪氨酸 (pY) 的肽通常通过抗体和/或基于金属亲和力的富集从细胞裂解物中分离，然后通过质谱法进行鉴定。在此，我们描述了磷酸酪氨酸肽的稳健两阶段分离和磷酸位点的质谱辅助鉴定，以表征未刺激的非小细胞肺癌（NSCLC）细胞系中的基础信号网络。
Robust isolation and identification of peptides phosphorylated at their tyrosine residues are key steps in deciphering complex signaling networks governed by protein tyrosine kinases, including kinases involved in oncogenesis. Phosphotyrosine (pY)-containing peptides are commonly isolated from cellular lysates by means of antibody and/or metal affinity-based enrichment followed by their identification by mass spectrometry. Herein, we describe robust two-stage isolation of phosphotyrosine peptides and mass spectrometry-aided identification of phosphosites to characterize basal signaling networks in unstimulated non-small cell lung cancer (NSCLC) cell lines.