Palmitoylation targets the Calcineurin phosphatase to the Phosphatidylinositol 4-kinase complex at the plasma membrane

This dataset is associated with the publication Ulengin-Talkish, I. et. al., 2021. This manuscript is currently under revision in Nature Communications.
In this study, we elucidate a new signaling pathway regulated by the Ca2+-activated phosphatase, calcineurin (CN) by examining the understudied isoform CNAB1. Using multiple cell based biochemical assays including pulse-chase experiments with metabolic analog of palmitate and click chemistry, we show that CNAB1 is dynamically palmitoylated at two sites unique to its C-terminal tail. This dynamic palmitoylation, regulated by the ABHD17A depalmitoylase, confers unique localization to the plasma membrane (PM) and the golgi, substrate specificity and regulation to this isoform. We further uncover the PM-associated phosphatidylinositol 4-kinase complex to be a regulatory target of this isoform through which CN regulates PI4P production at the PM during signaling from the type-3 muscarinic receptor.
This dataset specifically contains indirect immunofluorescence experiments done in COS-7 cells, which were used for the analysis shown in figures 1, 2 and supplementary figure 1. These multichannel images were acquired on a single z-plane on LionheartTM FX automated widefield microscope with a 20X Plan Fluorite WD 6.6 NP 0.45 objective.

本数据集关联Ulengin-Talkish I. 等人2021年的研究成果，该稿件目前正在《Nature Communications》（自然·通讯）期刊进行审稿修订。
本研究聚焦此前未被充分探索的Ca²⁺激活的磷酸酶钙调神经磷酸酶（calcineurin, CN）同工型CNAB1，阐明了一条由其调控的全新信号通路。研究采用多种基于细胞的生化实验手段，包括使用棕榈酸代谢类似物的脉冲追踪实验与点击化学技术，证实CNAB1在其C端尾段特有的两个位点上发生动态棕榈酰化修饰。
这种由ABHD17A去棕榈酰化酶调控的动态棕榈酰化修饰，赋予该同工型独特的细胞膜（plasma membrane, PM）与高尔基体定位、底物特异性及调控特性。
研究进一步发现，与细胞膜结合的磷脂酰肌醇4-激酶复合物是该同工型的调控靶点，钙调神经磷酸酶可通过该复合物在3型毒蕈碱受体介导的信号转导过程中，调控细胞膜上的PI4P（磷脂酰肌醇4-磷酸）生成。
本数据集专属收录了在COS-7细胞中开展的间接免疫荧光实验，该实验用于分析图1、图2及补充图1中的结果。上述多通道图像均在单Z平面下，通过搭载20X Plan Fluorite WD 6.6 NP 0.45物镜的Lionheart™ FX自动化宽场显微镜采集获得。