Identification and mapping of central pair proteins by proteomic analysis.
Identification and mapping of central pair proteins by proteomic analysis.
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DOI：
10.2142/biophysico.bsj-2019048
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发表时间：
2020
影响因子：
1.5
通讯作者：
Huy Bui K
中科院分区：
文献类型：
--
作者：
Dai D;Ichikawa M;Peri K;Rebinsky R;Huy Bui K
真核生物的纤毛或鞭毛是微小的毛发状结构，是单细胞运动必不可少的，在哺乳动物的生物过程中起着重要作用。纤毛或鞭毛由九个双线微管围绕一对称为中心对（CP）的单线微管组成。总之，这种排列形成了一个典型的和高度保守的9+2轴突结构。纤毛是纤毛特有的一种结构，由一对结构二态的单线态微管组成，微管上装饰着许多相关蛋白。cp相关蛋白的突变引起几种不同的身体症状，称为纤毛病。因此，了解CP的结构是至关重要的。然而，人们对CP的蛋白质组成知之甚少。这是因为传统的CP蛋白鉴定方法大多受到现有的CP蛋白突变体衣藻的限制。近年来，基于质谱分析结果提出了更多的CP候选蛋白，但这些蛋白大多没有得到验证。在这项研究中，我们通过进行类似的全面CP蛋白质组分析，比较了衣藻菌株携带和不携带CP复合物的轴质体样品的质谱分析结果，重新评估了CP蛋白。我们发现了一组相似的CP候选蛋白和另外11个新的CP候选蛋白。此外，通过使用缺乏特异性CP亚结构的衣藻菌株，我们提出了一个更完整的CP蛋白定位模型。本研究为进一步了解CP复合物的功能奠定了新的基础。
Cilia or flagella of eukaryotes are small micro-hair like structures that are indispensable to single-cell motility and play an important role in mammalian biological processes. Cilia or flagella are composed of nine doublet microtubules surrounding a pair of singlet microtubules called the central pair (CP). Together, this arrangement forms a canonical and highly conserved 9+2 axonemal structure. The CP, which is a unique structure exclusive to motile cilia, is a pair of structurally dimorphic singlet microtubules decorated with numerous associated proteins. Mutations of CP-associated proteins cause several different physical symptoms termed as ciliopathies. Thus, it is crucial to understand the architecture of the CP. However, the protein composition of the CP was poorly understood. This was because the traditional method of identification of CP proteins was mostly limited by available Chlamydomonas mutants of CP proteins. Recently, more CP protein candidates were presented based on mass spectrometry results, but most of these proteins were not validated. In this study, we re-evaluated the CP proteins by conducting a similar comprehensive CP proteome analysis comparing the mass spectrometry results of the axoneme sample prepared from Chlamydomonas strains with and without CP complex. We identified a similar set of CP protein candidates and additional new 11 CP protein candidates. Furthermore, by using Chlamydomonas strains lacking specific CP sub-structures, we present a more complete model of localization for these CP proteins. This work has established a new foundation for understanding the function of the CP complex in future studies.