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

研究了绿藻衣藻reinhardtü中通道视紫红质（ChR1和ChR2）在体内和在动物细胞中表达后的功能特征。它们的n端一半构成直接光门控阳离子通道，作为光操纵神经元活动的工具越来越多地使用。[这部分项目不建议获得资助：对于两种通道视紫红质，c端一半的功能是完全未知的。]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发现的ChR c端磷酸化位点的作用。在衣藻reinhardtü基因组中发现了具有预测环化酶结构域的新rhodopslns。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已经研究好的环化酶的相应结构域，以表征预测结构域的功能。或者，将建立的环化酶结构域与预测的视紫红质环化酶结构域交换，以测试环化酶活性的光调制。

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.