Proteomic profiling of over 13,000 proteins across ten mouse tissues using multi-dimensional peptide pre-fractionation

Proteome-wide abundance profiling across tissues can help to provide insight into the biological mechanisms underlying tissue-specific function and potential related dysfunction and amelioration thereof. Here, we use sample multiplexing to profile the proteomes of ten diverse mouse tissues using TMTpro-based quantitative mass spectrometry. Our optimized workflow, incorporating two-dimensional peptide pre-fractionation (which included both basic pH reversed-phase and strong ion exchange chromatography) enabled the quantification of over 13,000 proteins across brain, brown fat, heart, kidney, liver, lung, skeletal muscle, spleen, ovaries, and testes. Global analysis revealed distinct proteome profiles for each tissue, with clear clustering patterns reflecting functional similarities and differences. We highlighted the abundance of numerous tissue-specific proteins, exemplified by Synapsin-1 in brain, Uncoupling protein 1 in brown fat, and Zona pellucida proteins in reproductive tissues. Gene ontologies and pathway analyses of the most relatively abundant proteins in each tissue revealed enrichment patterns consistent with known physiological functions. For instance, brain tissue showed enrichment for synaptic components and neurotransmission processes, while liver tissue was enriched for metabolic pathways. This dataset serves as a valuable resource for mapping tissue-specific protein landscapes in mammals, offering potential insights into the molecular mechanisms of tissue function.