Characterization of proteins involved in the regulation of sperm motility and their protein network

参与精子活力调节的蛋白质及其蛋白质网络的表征

• 批准号: 15570061
• 负责人: INABA Kazuo
• 金额: $ 2.37万
• 依托单位: University of Tsukuba (2004)Tohoku University (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570061/
• 关键词: ascidians; sperm; flagellar motility; dynein; axonemes; radial spoke; HSP40; leucine-rich repeat; タンパク質リン酸化; プロテオミクス; 14-3-3タンパク質; 分子シャペロン

The research was carried out to elucidate the molecular network or cascade from the reception of signaling molecule to the activation of motile machinery in sperm cell. As a result a part of the proteins involved in this signaling pathway have been identified and the molecular interactions among the molecules have been revealed. In the research period twelve proteins that undergo changes in electric point or in the amount were detected in the sperm of the ascidian Ciona intestinalis on two-dimentional gel electrophoresis and peptide mass fingerprinting, Among them, potentially important axonemal proteins could be identified : outer arm dynein intermediate chain2(IC2), an intermediate chain of inner arm dynein termed IC116, radial spoke component LRR37, the regulatory subunit of cAMP-dependent protein kinase and a 94 kDa novel armadillo repeat protein, Another proteins are though to be involved in the reception of signals or intracellular signal transduction, such as SP17-like protein, TER ATPasae,TEFB,and 14-3-3 proteins. On the other hand, protein-protein interaction was examined by means of selective extraction of the axonemes, a series of columm chromatography and immunoprecipitation. As a result, a subcomplex constitution the radial spoke could be isolated. The subcomptex was composed of AKAP-like protein (RSP3), HSP40, two MORN repeat proteins, RSP4/6 and several novel proteins whose homologs are also present in human but the functions have been unrevealed. The molecular interaction in this subcomplex would shed light on the molecular function of radial spoke and ultimately on the mechanism of flagellar motility conserved from ascidian to human. The result also suggests that molecular chaperone is involved in the formation, maintenance or regulation of flagellar motility.

进行了研究以阐明从信号分子的接收到精子细胞中通气机械的激活的分子网络或级联。结果，已经鉴定出参与该信号通路的一部分蛋白质，并揭示了分子之间的分子相互作用。在研究期间，在二维凝胶电泳和肽质量指纹指纹的Xiona intestinalis的精子中检测到了十二个蛋白质或数量发生变化的蛋白质，其中可能会鉴定出潜在的重要轴突蛋白，可以鉴定出潜在的重要轴突蛋白：外臂dynein Intremidied chinimed Chain 2（IC2）ININDIRED SINDRIDIAL STERMIADE IC2 IC2 IC2 IC2 IC2，IC2 component LRR37, the regulatory subunit of cAMP-dependent protein kinase and a 94 kDa novel armadillo repeat protein, Another proteins are though to be involved in the reception of signals or intracellular signal transduction, such as SP17-like protein, TER ATPasae,TEFB,and 14-3-3 proteins.另一方面，通过选择性提取轴突，一系列横向色谱和免疫沉淀来检查蛋白质 - 蛋白质相互作用。结果，可以隔离径向辐条的子复合体。子成棒由AKAP样蛋白（RSP3），HSP40，两个早晨重复蛋白，RSP4/6和几种新型蛋白质组成，它们的同源物也存在于人类中，但功能已不明式。该子复合物中的分子相互作用将阐明径向的分子功能，并最终介绍了从海藻到人的鞭毛运动机理。结果还表明，分子伴侣参与了鞭毛运动的形成，维持或调节。

项目成果

Inaba, K.et al.: "Control of flatfish sperm motility by CO_2 and carbonic anhydrase"Cell Motil Cytoskeleton. 55. 174-187 (2003)

Inaba, K.等人：“CO_2 和碳酸酐酶对比目鱼精子活力的控制”Cell Motil 细胞骨架。

Control of flatfish sperm motility by CO2 and carbonic anhydrase

• DOI: 10.1002/cm.10119
• 发表时间: 2003-07-01
• 期刊: CELL MOTILITY AND THE CYTOSKELETON
• 作者: Inaba, K;Dréanno, C;Cosson, J
• 通讯作者: Cosson, J

DC3, the 21-kD subunit of the outer dynein arm-docking complex (ODA-DC), is a novel EF-hand protein important for assembly of both the outer arm and the ODA-DC

DC3 是外动力蛋白臂对接复合物 (ODA-DC) 的 21 kD 亚基，是一种新型 EF 手蛋白，对于外臂和 ODA-DC 的组装非常重要

• 发表时间: 2003
• 期刊: Mol.Biol.Cell 14
• 作者: Casey;DM.et al.
• 通讯作者: DM.et al.

Padma, P.et al.: "Identification of a novel leucine-rich repeat as a component of flagellar radial spoke in the ascidian Ciona intestinalis"Mol.Biol.Cell. 14. 774-785 (2003)

Padma, P. 等人：“鉴定一种新的富含亮氨酸的重复序列，作为海鞘海鞘鞭毛径向辐条的组成部分”Mol.Biol.Cell。

Inaba, K.: "Molecular architecture of sperm flagella : molecules for motility and signaling"Zool.Sci.. 20. 1043-1056 (2003)

Inaba, K.：“精子鞭毛的分子结构：运动和信号传导分子”Zool.Sci.. 20. 1043-1056 (2003)