A novel inducible retroviral expression system for StrepHA-tandem affinity purification mass-spectrometry-based proteomics

Tandem-affinity purification mass spectrometry using the streptavidin-hemagglutinin (SH)-tag has been successfully employed to map signaling networks in several large-scale studies. However its application has so far been restricted to the small number of Flp/FRT-recombination competent cell lines. We present pRSHIC, a novel retroviral, doxycycline-inducible Tet-On vector system suitable for expression of SH-tagged target proteins in a wide range of human and mouse cell systems. The additional feature of concomitant reporter fluorophore expression makes pRSHIC a valuable tool for a diverse set of phenotypic analyses beyond TAP-MS experiments. The dataset demonstrates the application of pRSHIC for TAP-MS analysis of two showcase bait proteins involved in cancer cell proliferation as well as cell death induction and identified novel high- confidence interacting proteins with possible pharmacological intervention potential.