DiPAK

(from paper) DiPAK consists of two AK2(115)fluorescent protein fusions. The 'sensor unit' harbours the first 15 amino acids of AK2, which can be sequentially processed by MAP and DPP8/9 triggering proteasomal degradation of the sensor. The normalizer unit contains the respective S4P variant of AK2, which is insensitive to DPP8/9 processing, thus rendering the normalizer levels independent of DPP8/9 activity. Sensor and normalizer signals can be analysed based on their fluorescence properties (mEGFP or mKate2 fluorescence) or via detection of the protein tags (Strep and HA). The genes encoding both fusion proteins are combined on one plasmid and separated by an internal ribosomal entry site allowing for sensor and normalizer expression in a fixed ratio. The normalized DiPAK ratio is calculated by dividing the sensor by the normalizer signals and inversely correlates to DPP8/9 activity