Stable isotope labeling and label-free proteomics of Drosophila parkin null mutants.
Stable isotope labeling and label-free proteomics of Drosophila parkin null mutants.
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DOI：
10.1021/pr9006238
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发表时间：
2009-10
影响因子：
4.4
通讯作者：
Clemmer, David E.
中科院分区：
文献类型：
--
作者：
Xun, Zhiyin;Kaufman, Thomas C.;Clemmer, David E.
帕金森病（PD）的特征是黑质多巴胺能神经元的丧失和胞浆内路易体（LBs）的形成。编码E3泛素蛋白连接酶的帕金森氏蛋白的功能丧失突变是家族性帕金森氏病（常染色体隐性遗传性少年帕金森氏症）的主要原因。果蝇parkin无突变体表现为肌肉变性和线粒体功能障碍，为研究帕金森病相关分子通路提供了动物模型。为了确定Parkin发病机制中涉及的蛋白质改变，我们利用全球内标准技术（GIST）和提取离子色谱峰面积（XICPA）无标记方法对果蝇Parkin无突变体和年龄匹配对照进行了定量蛋白质组学分析。在两个独立的生物重复中，通过结合XICPA和GIST测量，至少鉴定了两个肽段，共对375个蛋白质进行了定量。16个蛋白表现出显著的改变。失调蛋白中有7种与能量代谢有关，其中6种下调。其中，血清蛋白1α、血清蛋白1β、血清蛋白1γ和脂肪体蛋白1的表达量上调了10倍以上，其中脂肪体蛋白1的表达量明显升高。这些发现提示能量代谢和蛋白质转运蛋白活性途径的异常可能与帕金森相关性ARJP的发病机制有关。
Parkinson’s disease (PD) is characterized by loss of dopaminergic neurons in the substantia nigra and formation of intracytoplasmic Lewy bodies (LBs). Loss-of-function mutations in parkin which encodes an E3 ubiqutin protein ligase contribute to a predominant cause of a familial form of PD termed autosomal recessive juvenile Parkinsonism (AR-JP). Drosophila parkin null mutants display muscle degeneration and mitochondrial dysfunction, providing an animal model to study Parkin-associated molecular pathways in PD. To define protein alterations involved in Parkin pathogenesis, we performed quantitative proteomic analyses of Drosophila parkin null mutants and age-matched controls utilizing both global internal standard technology (GIST) and extracted ion chromatogram peak area (XICPA) label-free approaches. A total of 375 proteins were quantified with a minimum of two peptide identifications from the combination of the XICPA and GIST measurements applied to two independent biological replicates. Sixteen proteins exhibited significant alteration. Seven of the dysregulated proteins are involved in energy metabolism, of which six were down-regulated. All five proteins involved in transporter activity exhibited higher levels, of which larval serum protein 1α, larval serum protein 1β, larval serum protein 1γ, and fat body protein 1 showed > 10-fold up-regulation and substantially higher level of fat body protein 1 was confirmed by Western blot analysis. These findings suggest that abnormalities in energy metabolism and protein transporter activity pathways may be associated with the pathogenesis of Parkin-associated ARJP.
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影响因子：
11.8
作者：
Arama, E;Agapite, J;Steller, H
通讯作者：
Steller, H
影响因子：
4.8
作者：
Goldberg, MS;Fleming, SM;Shen, J
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Shen, J
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3.1
作者：
Haywood, Annika F. M.;Staveley, Brian E.
通讯作者：
Staveley, Brian E.
影响因子：
14.9
作者：
LIMBACH, KJ;WU, R
通讯作者：
WU, R
影响因子：
11.4
作者：
Arama, E;Bader, M;Steller, H
通讯作者：
Steller, H