A Method for Rapid Differential Protein Expression Profiling from Tissue Using 'Shotgun-based' LC-MS/MS and Spectral Counting

摘要

The reported proteomics-based approach provides a rapid procedure to profile differential protein expression and posttranslational modifications from tissue with high sensitivity and dynamic range of quantitation. A small amount of animal tissue (<5 mg) is needed for this method allowing for detailed proteomic analysis of heterogeneous tissues such as the brain in which differential expression of certain proteins may vary significantly across different regions in response to a specific experimental treatment or pathophysiological state. Successful application of this method to profile estrogen-regulated protein expression in mouse brain and uterus was demonstrated. Taken together, our results demonstrate the utility of a gel-free, mass spectrometry-based approach to rapidly screen for differentially expressed proteins from tissue, ultimately providing a strong analytical foundation for numerous applications ranging from high-throughput drug discovery and development, toxicological screening, and characterization of pathological tissue specimens.