Molecular characterization of radial spoke subcomplex containing radial spoke protein 3 and heat shock protein 40 in sperm flagella of the ascidian Ciona intestinalis.
Molecular characterization of radial spoke subcomplex containing radial spoke protein 3 and heat shock protein 40 in sperm flagella of the ascidian Ciona intestinalis.
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DOI：
10.1091/mbc.e04-09-0784
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发表时间：
2004-11
影响因子：
3.3
通讯作者：
Yuhkoh Satouh;P. Padma;T. Toda;N. Satoh;H. Ide;K. Inaba
中科院分区：
文献类型：
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作者：
Yuhkoh Satouh;P. Padma;T. Toda;N. Satoh;H. Ide;K. Inaba
热休克蛋白(HSP)40的成员调节HSP70蛋白的折叠活性，并帮助该分子伴侣系统在各种类型的细胞事件中实现功能专门化。我们最近在海鞘中发现Hsp40是鞭毛轴丝的一种成分，提示Hsp40相关的伴侣系统与鞭毛功能之间存在相关性。在本研究中，我们发现，用0.5MKI溶液从KCl3处理的轴丝中提取Ciona37-kDa Hsp40，并通过凝胶过滤和离子交换层析，与放射状辐射蛋白(RSP)3和几种蛋白质形成复合体。用基质辅助激光解吸电离/飞行时间/质谱仪进行多肽指纹图谱分析表明，该复合体中的其他蛋白质包括一个海胆鱼头蛋白的同源蛋白(RSP4/6的同源蛋白)、一个与meichroacidin序列相似的膜占据和识别连接点重复蛋白以及一个功能未知的33 kDa蛋白。该复合体中不包括辐条头部蛋白LRR37，表明该复合体构建了径向辐条的茎。免疫电子显微镜显示，Hsp40定位于轮辐柄的远端，可能位于轮辐头与柄的交界处。
Members of the heat-shock protein (HSP)40 regulate the protein folding activity of HSP70 proteins and help the functional specialization of this molecular chaperone system in various types of cellular events. We have recently identified Hsp40 as a component of flagellar axoneme in the ascidian Ciona intestinalis, suggesting a correlation between Hsp40 related chaperone system and flagellar function. In this study, we have found that Ciona 37-kDa Hsp40 is extracted from KCl-treated axonemes with 0.5 M KI solution and comigrates with radial spoke protein (RSP)3 along with several proteins as a complex through gel filtration and ion exchange columns. Peptide mass fingerprinting with matrix-assisted laser desorption ionization/time of flight/mass spectrometry revealed that other proteins in the complex include a homolog of sea urchin spokehead protein (homolog of RSP4/6), a membrane occupation and recognition nexus repeat protein with sequence similarity with meichroacidin, and a functionally unknown 33-kDa protein. A spoke head protein, LRR37, is not included in the complex, suggesting that the complex constructs the stalk of radial spoke. Immunoelectron microscopy indicates that Hsp40 is localized in the distal portion of spoke stalk, possibly at the junction between spoke head and the stalk.