Mass spectrometric analysis of microtubule co‐sedimented proteins from rat brain
Mass spectrometric analysis of microtubule co‐sedimented proteins from rat brain
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DOI：
10.1111/j.1365-2443.2008.01175.x
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发表时间：
2008-04
期刊：
影响因子：
2.1
通讯作者：
Tatsuhiko Sakamoto;Akiyoshi Uezu;S. Kawauchi;T. Kuramoto;K. Makino;Kazuaki Umeda;N. Araki;Hideo Baba;H. Nakanishi
中科院分区：
文献类型：
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作者：
Tatsuhiko Sakamoto;Akiyoshi Uezu;S. Kawauchi;T. Kuramoto;K. Makino;Kazuaki Umeda;N. Araki;Hideo Baba;H. Nakanishi
微管（MT）在多种细胞功能中发挥着至关重要的作用，例如有丝分裂、囊泡运输和细胞运动。MT还组成专门的结构，如中心体，纺锤体和纤毛。然而，这些基于MT的功能和结构的分子机制尚未完全理解。在这里，我们分析了MT共沉淀蛋白质从大鼠脑串联质谱（MS）离子交换柱层析。共鉴定出391种蛋白质。这些蛋白质分为12类：57种MT细胞骨架蛋白，包括MT相关蛋白（MAPs）和马达蛋白; 66种其他细胞骨架蛋白; 4种中心体蛋白; 10种伴侣蛋白; 5种高尔基体蛋白; 7种线粒体蛋白; 62种核酸结合蛋白; 14种核蛋白; 13种核糖体蛋白; 28种囊泡转运蛋白; 83个具有不同功能和/或定位的蛋白质;和42个未表征的蛋白质。在这些未知的蛋白质中，有6种蛋白质在培养的细胞中表达，从而鉴定出中心体和纤毛的三种新成分。我们目前的方法并不特异于MAP，但可用于鉴定低丰度的新型MAP和基于MT的结构的组分。我们的分析为未来研究MT功能和结构的分子机制提供了广泛的潜在候选人名单。
Microtubules (MTs) play crucial roles in a variety of cell functions, such as mitosis, vesicle transport and cell motility. MTs also compose specialized structures, such as centrosomes, spindles and cilia. However, molecular mechanisms of these MT‐based functions and structures are not fully understood. Here, we analyzed MT co‐sedimented proteins from rat brain by tandem mass spectrometry (MS) upon ion exchange column chromatography. We identified a total of 391 proteins. These proteins were grouped into 12 categories: 57 MT cytoskeletal proteins, including MT‐associated proteins (MAPs) and motor proteins; 66 other cytoskeletal proteins; 4 centrosomal proteins; 10 chaperons; 5 Golgi proteins; 7 mitochondrial proteins; 62 nucleic acid‐binding proteins; 14 nuclear proteins; 13 ribosomal proteins; 28 vesicle transport proteins; 83 proteins with diverse function and/or localization; and 42 uncharacterized proteins. Of these uncharacterized proteins, six proteins were expressed in cultured cells, resulting in the identification of three novel components of centrosomes and cilia. Our present method is not specific for MAPs, but is useful for identifying low abundant novel MAPs and components of MT‐based structures. Our analysis provides an extensive list of potential candidates for future study of the molecular mechanisms of MT‐based functions and structures.