The function of the C-terminal half of Channelrhodopsins and of predicted novel rhodopsins from the green alga Chlamydomonas reinhardtii

通道视紫红质的 C 端一半的功能和来自绿藻莱茵衣藻的预测新型视紫红质的功能

• 批准号: 146946707
• 负责人: Professor Dr. Georg Nagel
• 金额: --
• 依托单位: Lehrstuhl für Physiologie II - Neurophysiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/146946707?language=en
• 关键词: function; terminal; half; Channelrhodopsins; predicted

Channelrhodopsins (ChR1 and ChR2) from the green alga Chlamydomonas reinhardtü were functionally characterized in-vivo and after expression In animal cells. Their N-terminal half constitutes a directly light-gated cation channel which finds increasing use as a tool to light-manipulate neuronal activity. [This part of the project was not recommended for funding: The function of the C-terminal half is completely unknown for both channelrhodopsins. We plan to investigate the role of the recently identified phosphorylation sites on the C-terminus of ChR by analyzing functional consequences of co-expressing suitable kinases or by expressing phosphorylation site mutants.] New rhodopslns with predicted cyclase domains were identified in the genome of Chlamydomonas reinhardtü. We plan to express the predicted proteins or domains derived from these sequences. If functional expression of the whole protein fails we plan to swap domains with predicted cyclase activity into the corresponding domains of well studied cyclases to characterize the function of the predicted domain. Alternatively, established cyclase domains will be swapped against the predicted cyclase domain of the rhodopsin to test for light-modulation of cyclase activity.

对来自绿色莱茵衣藻的视紫红质（ChR 1和ChR 2）在体内和在动物细胞中表达后进行功能表征。它们的N-末端的一半构成了一个直接的光门控阳离子通道，发现越来越多的使用作为一种工具，光操纵神经元的活动。[This该项目的一部分不建议资助：两种通道视紫红质的C-末端一半的功能完全未知。我们计划通过分析共表达合适激酶或表达磷酸化位点突变体的功能后果来研究最近鉴定的ChR C末端磷酸化位点的作用。在莱茵衣藻基因组中鉴定出具有预测环化酶结构域的新的视紫红质。我们计划表达来自这些序列的预测蛋白或结构域。如果整个蛋白质的功能表达失败，我们计划将具有预测环化酶活性的结构域交换到充分研究的环化酶的相应结构域中，以表征预测结构域的功能。或者，将建立的环化酶结构域与视紫红质的预测环化酶结构域交换，以测试环化酶活性的光调节。