Add reference The present dataset provides the first description of Pseudomonas aeruginosa inner membrane proteome. To achieve this, we combined efficient separation of membranes from PAO1 reference strain using discontinuous sucrose gradient centrifugation and mass spectrometry-based proteomic analysis. Three biological replicates were prepared and analysed twice by nanoLC-MS/MS (LTQ-Orbitrap Velos pro). Using Mascot (v.2.4), IRMa (v.1.30.4) and homemade hEIDI (v.1.13), a core list of 991 non-redundant proteins was established.