Phosphoproteome of human glioblastoma initiating cells reveals novel signaling regulators encoded by the transcriptome.
Phosphoproteome of human glioblastoma initiating cells reveals novel signaling regulators encoded by the transcriptome.
复制标题
DOI：
10.1371/journal.pone.0043398
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Oyama M
中科院分区：
文献类型：
--
作者：
Kozuka-Hata H;Nasu-Nishimura Y;Koyama-Nasu R;Ao-Kondo H;Tsumoto K;Akiyama T;Oyama M
胶质母细胞瘤是最具侵袭性的肿瘤之一，预后差。尽管到目前为止已经进行了各种研究，但还没有针对胶质母细胞瘤患者的有效治疗方法。为了系统地阐明恶性脑肿瘤中激活的异常信号机制，我们利用高分辨率纳米流LC-MS/MS系统，结合SILAC技术，研究了胶质母细胞瘤起始细胞的磷酸化蛋白质组动力学。通过二氧化钛小球富集磷酸肽，基于碰撞诱导解离和高能C-TRAP解离两种解离方法，从2,282个磷酸化蛋白质中鉴定出6,073个磷酸肽。基于SILAC的定量描述了表皮生长因子刺激时上调的516个磷酸化位点和下调的275个磷酸化位点，包括干细胞标记物上的磷酸化位点的综合状态，如巢蛋白。非常有趣的是，我们深入的磷酸蛋白质组分析导致了由人类转录本的未定义序列区域编码的新的磷酸化分子的鉴定，其中一个受外部刺激调节的人胶质母细胞瘤启动细胞。我们的结果揭示了由人类转录组定义的调节性磷蛋白质组的扩大的多样性。
Glioblastoma is one of the most aggressive tumors with poor prognosis. Although various studies have been performed so far, there are not effective treatments for patients with glioblastoma. In order to systematically elucidate the aberrant signaling machinery activated in this malignant brain tumor, we investigated phosphoproteome dynamics of glioblastoma initiating cells using high-resolution nanoflow LC-MS/MS system in combination with SILAC technology. Through phosphopeptide enrichment by titanium dioxide beads, a total of 6,073 phosphopeptides from 2,282 phosphorylated proteins were identified based on the two peptide fragmentation methodologies of collision induced dissociation and higher-energy C-trap dissociation. The SILAC-based quantification described 516 up-regulated and 275 down-regulated phosphorylation sites upon epidermal growth factor stimulation, including the comprehensive status of the phosphorylation sites on stem cell markers such as nestin. Very intriguingly, our in-depth phosphoproteome analysis led to identification of novel phosphorylated molecules encoded by the undefined sequence regions of the human transcripts, one of which was regulated upon external stimulation in human glioblastoma initiating cells. Our result unveils an expanded diversity of the regulatory phosphoproteome defined by the human transcriptome.
登录
查看更多内容
影响因子：
4.4
作者：
Nilsson, Carol L.;Dillon, Roslyn;Conrad, Charles A.
通讯作者：
Conrad, Charles A.
影响因子：
10.3
作者：
Legler, JM;Ries, LAG;Linet, MS
通讯作者：
Linet, MS
影响因子：
7
作者：
Olsen, JV;de Godoy, LMF;Mann, M
通讯作者：
Mann, M
影响因子：
5.3
作者：
Murakami, M;Ichisaka, T;Yamanaka, S
通讯作者：
Yamanaka, S
影响因子：
50.3
作者：
Lee, Jeongwu;Kotliarova, Svetlana;Fine, Howard A.
通讯作者：
Fine, Howard A.