Phosphoproteomic analysis reveals site-specific changes in GFAP and NDRG2 phosphorylation in frontotemporal lobar degeneration.
Phosphoproteomic analysis reveals site-specific changes in GFAP and NDRG2 phosphorylation in frontotemporal lobar degeneration.
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DOI：
10.1021/pr100666c
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发表时间：
2010-12-03
影响因子：
4.4
通讯作者：
Levey, Allan I.
中科院分区：
文献类型：
--
作者：
Herskowitz, Jeremy H.;Seyfried, Nicholas T.;Duong, Duc M.;Xia, Qiangwei;Rees, Howard D.;Gearing, Marla;Peng, Junmin;Lah, James J.;Levey, Allan I.
关键词：
额颞叶变性（FTLD）是一种进行性神经退行性疾病，其特征在于行为异常、人格改变、语言功能障碍，并且可以与运动神经元疾病的发展同时发生。FTLD的一种主要病理形式的特征在于泛素化和磷酸化的TAR DNA结合蛋白-43（TDP-43）的细胞内沉积，这表明磷酸化事件中的失调可能有助于疾病进展。然而，迄今为止，系统分析的磷酸蛋白质组在FTLD大脑还没有报道。在这项研究中，我们采用固定化金属亲和色谱（IMAC），然后液相色谱-串联质谱（LC-MS/MS），以确定磷酸肽从FTLD和年龄匹配的控制死后人脑组织。使用这种方法，我们确定了786磷酸肽在额叶皮层（控制和FTLD），其中磷酸肽的人口约占50%的总肽分析。使用光谱计数的无标记定量揭示了FTLD磷酸化蛋白质组中具有显著变化的六种蛋白质。在FTLD中，N-myc下游调节基因2（NDRG 2）和胶质细胞酸性蛋白（GFAP）的磷酸化谱数目增加，而微管相关蛋白1A（MAP 1A）、网状蛋白4（reticulon 4（RTN4;也称为神经突生长抑制剂（Nogo）），蛋白激酶C γ（PRKCG），和热休克蛋白90kDa α，A类成员1（HSP 90 AA 1）与对照脑相比具有显著更少的磷酸光谱。为了验证这些差异，我们通过免疫印迹分析检测了FTLD脑中NDRG2的磷酸化，并使用磷酸丝氨酸-13（pSer13）GFAP单克隆抗体，我们通过免疫印迹显示FTLD病例中pSer13 GFAP水平的增加伴随着总体GFAP水平的增加。这些数据突出了结合蛋白质组学和磷酸化蛋白质组学策略来表征死后人脑组织的效用。
Frontotemporal lobar degeneration (FTLD) is a progressive neurodegenerative disease characterized by behavioral abnormalities, personality changes, language dysfunction, and can co-occur with the development of motor neuron disease. One major pathological form of FTLD is characterized by intracellular deposition of ubiquitinated and phosphorylated TAR DNA binding protein-43 (TDP-43), suggesting that dysregulation in phosphorylation events may contribute to disease progression. However, to date systematic analysis of the phosphoproteome in FTLD brains has not been reported. In this study we employed immobilized metal affinity chromatography (IMAC) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify phosphopeptides from FTLD and age-matched control postmortem human brain tissue. Using this approach we identified 786 phosphopeptides in frontal cortex (control and FTLD), in which the population of phosphopeptides represented approximately 50% of the total peptides analyzed. Label free quantification using spectral counts revealed six proteins with significant changes in the FTLD phosphoproteome. N-myc-downstream regulated gene 2 (NDRG2) and glial fibrillary acidic protein (GFAP) had an increased number of phosphospectra in FTLD, whereas microtubule associated protein 1A (MAP1A), reticulon 4 (RTN4; also referred to as neurite outgrowth inhibitor (Nogo)), protein kinase C gamma (PRKCG), and heat shock protein 90kDa alpha, class A member 1(HSP90AA1) had significantly fewer phosphospectra compared to control brain. To validate these differences, we examined NDRG2 phosphorylation in FTLD brain by immunoblot analyses, and using a phosphoserine-13 (pSer13) GFAP monoclonal antibody we show an increase in pSer13 GFAP levels by immunoblot concomitant with increased overall GFAP levels in FTLD cases. These data highlight the utility of combining proteomic and phosphoproteomic strategies to characterize postmortem human brain tissue.
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影响因子：
64.5
作者：
ABELIOVICH, A;CHEN, C;TONEGAWA, S
通讯作者：
TONEGAWA, S
DOI：
10.1073/pnas.94.1.298
发表时间：
1997-01-07
影响因子：
11.1
作者：
Alonso, AD;GrundkeIqbal, I;Iqbal, K
通讯作者：
Iqbal, K
影响因子：
6
作者：
Cairns, Nigel J.;Neumann, Manuela;Mackenzie, Ian R. A.
通讯作者：
Mackenzie, Ian R. A.
影响因子：
46.9
作者：
Ficarro, SB;McCleland, ML;White, FM
通讯作者：
White, FM
影响因子：
6.2
作者：
BEACH, TG;WALKER, R;MCGEER, EG
通讯作者：
MCGEER, EG