Deep proteome mapping of mouse kidney based on OFFGel prefractionation reveals remarkable protein post- translational modifications.
Deep proteome mapping of mouse kidney based on OFFGel prefractionation reveals remarkable protein post- translational modifications.
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DOI：
10.1021/pr401122m
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发表时间：
2014-03-07
影响因子：
4.4
通讯作者：
Yamamoto, Tadashi
中科院分区：
文献类型：
--
作者：
Magdeldin, Sameh;Yamamoto, Keiko;Yoshida, Yutaka;Xu, Bo;Zhang, Ying;Fujinaka, Hidehiko;Yaoita, Eishin;Yates, John R., III;Yamamoto, Tadashi
由于样本的复杂性和广泛的动态范围，进行全面的无偏倚蛋白质组分析是一个非同寻常的挑战，特别是在真核组织中。因此，在质谱分析之前进行的预分馏步骤对于减少复杂的生物基质并允许进行深入分析至关重要。在这里，我们演示了使用OFFGel预分馏来鉴定比非分馏样品中更多的低丰度和疏水性蛋白质。此外，肾脏蛋白质样本的OFFGel预分离能够通过检测PTM来揭示蛋白质功能相关性，特别是当预分离以有针对性的浓缩策略(如二氧化钛磷酸浓缩)增强时。本研究中使用的OFFGel-二氧化钛组合与其他全球磷酸蛋白质组学方法(SCX-二氧化钛、ERLIC-二氧化钛或HILIC-二氧化钛)相当。详细的小鼠肾脏蛋白质组和浓缩的磷酸肽为更好地了解肾脏蛋白修饰机制的工作机制提供了一个有用的平台，并最终将有助于我们了解病理过程和正常的生理肾功能。
Performing a comprehensive nonbiased proteome analysis is an extraordinary challenge due to sample complexity and wide dynamic range, especially in eukaryotic tissues. Thus, prefractionation steps conducted prior to mass spectrometric analysis are critically important to reduce complex biological matrices and allow in-depth analysis. Here we demonstrated the use of OFFGel prefractionation to identify more low abundant and hydrophobic proteins than in a nonfractionated sample. Moreover, OFFGel prefractionation of a kidney protein sample was able to unveil protein functional relevance by detecting PTMs, especially when prefractionation was augmented with a targeted enrichment strategy such as TiO2 phospho-enrichment. The OFFGel-TiO2 combination used in this study was comparable to other global phosphoproteomics approaches (SCX-TiO2, ERLIC-TiO2, or HILIC-TiO2). The detailed mouse kidney proteome with the phosphopeptide enrichment presented here serves as a useful platform for a better understanding of how the renal protein modification machinery works and, ultimately, will contribute to our understanding of pathological processes as well as normal physiological renal functions.
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影响因子：
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作者：
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通讯作者：
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作者：
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通讯作者：
Yamamoto, Tadashi