Ultrafast and Reproducible Proteomics from Small Amounts of Heart Tissue Enabled by Azo and timsTOF Pro

• DOI: 10.1021/acs.jproteome.1c00446
• Publication Date: 2021-07-08T19:22:25Z
• Authors: Timothy J. Aballo; David S. Roberts; Jake A. Melby; Kevin M. Buck; Kyle A. Brown; Ying Ge
• Keywords: Proteome; Sample Preparation; Bottom-up proteomics

Global bottom-up mass spectrometry (MS)-based proteomics is widely used for protein identification and quantification to achieve a comprehensive understanding of the composition, structure, function proteome. However, traditional sample preparation methods are time-consuming, typically including overnight tryptic digestion, extensive cleanup remove MS-incompatible surfactants, offline fractionation reduce proteome complexity prior online liquid chromatography-tandem (LC-MS/MS) analysis. Thus, there need fast, robust, reproducible method from complex proteomes. Herein, we developed an ultrafast enabled by Azo, photocleavable, MS-compatible surfactant that effectively solubilizes proteins promotes rapid combined with Bruker timsTOF Pro, which enables deeper coverage through trapped ion mobility (TIMS) parallel accumulation-serial fragmentation (PASEF) peptides. We applied this analyze human cardiac identified nearly 4000 groups as little 1 mg heart tissue in single one-dimensional LC-TIMS-MS/MS run high reproducibility. Overall, anticipate ultrafast, empowered both Azo Pro will be generally applicable greatly accelerate throughput large-scale quantitative proteomic studies. Raw data available via MassIVE repository identifier MSV000087476.