Native Mass Spectrometry Imaging and In Situ Top-Down Identification of Intact Proteins Directly from Tissue

Mass spectrometry imaging (MSI) provides information on the spatial distribution of molecules within a biological substrate without the requirement for labeling. Its broad specificity, i.e., the capability to spatially profile any analyte ion detected, constitutes a major advantage over other imaging techniques. A separate branch of mass spectrometry, native mass spectrometry, provides information relating to protein structure through retention of solution-phase interactions in the gas phase. Integration of MSI and native mass spectrometry (“native MSI”) affords opportunities for simultaneous acquisition of spatial and structural information on proteins directly from their physiological environment. Here, we demonstrate significant improvements in native MSI and associated protein identification of intact proteins and protein assemblies in thin sections of rat kidney by use of liquid extraction surface analysis on a state-of-the-art Orbitrap mass spectrometer optimized for intact protein analysis. Proteins of up to 47 kDa, including a trimeric protein complex, were imaged and identified.