Proteomic analysis of the eyespot of Chlamydomonas reinhardtii and functional characterization of components involved in tactic movements

莱茵衣藻眼斑的蛋白质组学分析和参与战术运动的成分的功能表征

• 批准号: 36330673
• 负责人: Professor Georg Kreimer
• 金额: --
• 依托单位: Lehrstuhl für Zellbiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/36330673?language=en
• 关键词: Proteomic; analysis; eyespot; Chlamydomonas; reinhardtii

The model green alga Chlamydomonas reinhardtii exhibits photo- and chemotactic movements. Both signaling pathways converge possibly at the level of membrane depolarization and Ca2+ is intricately involved in both responses at different levels. We have developed a procedure to purify the primitive visual system for phototaxis, the eyespot, of this alga. In the following proteomic approach, 202 proteins of the eyespot fraction were identified with at least two different peptides (984 in total). The identified proteins include e.g. structural components, photoreceptors, as well as members of metabolic and putative signaling pathways for tactic movements. Now our aim is to get functional information about two selected proteins from this proteome: (i) a Ca2+ sensing receptor and (ii) a protein related to a cyanobacterial chemotaxis receptor. Among the identified proteins were also kinases and phosphatases, indicating that phosphorylation/ dephosphorylation is important for signaling related to the eyespot. Silencing of one of the kinases, casein kinase 1(CK1), affects the circadian control of phototaxis. However, nothing is yet known about the targets of kinases in the eyespot. Therefore we will also analyze the eyespot phosphoproteome and do comparative quantitative phosphoproteomics with eyespots isolated from CK1-silenced and wild-type cells to identify potential targets of CK1 and other kinases in this primitive visual system. Thereby we hope to contribute to the further understanding of signaling pathways involved in tactic movements of C. reinhardtii.

模型绿色衣藻表现出光和趋化运动。这两种信号通路可能在膜去极化水平上会聚，并且Ca 2+在不同水平上复杂地参与这两种反应。我们已经开发了一个程序，以净化的原始视觉系统的趋光性，眼点，这条鱼。在以下蛋白质组学方法中，用至少两种不同的肽鉴定了眼斑组分的202种蛋白质（总共984种）。所鉴定的蛋白质包括例如结构组分、光感受器以及用于策略运动的代谢和推定信号传导途径的成员。现在，我们的目标是从这个蛋白质组中获得两个选定的蛋白质的功能信息：（i）Ca 2+感应受体和（ii）与蓝藻趋化性受体相关的蛋白质。在所鉴定的蛋白质中还包括激酶和磷酸酶，这表明磷酸化/去磷酸化对于与眼点相关的信号传导是重要的。其中一种激酶酪蛋白激酶1（CK 1）的沉默影响趋光性的昼夜节律控制。然而，目前还不清楚眼斑中激酶的靶点。因此，我们还将分析眼斑磷酸化蛋白质组，并与从CK 1沉默和野生型细胞中分离的眼斑进行比较定量磷酸化蛋白质组学，以确定CK 1和其他激酶在这个原始视觉系统中的潜在靶点。从而为进一步了解C.莱因哈德氏菌