Improved methodology for stable isotope labeling and LC-MS/MS analysis for proteomic scale analysis of protein turnover in plants

摘要

An increasing body of data documents that protein turnover is critical to the cellular regulatory processes that allow cells to respond rapidly to intracellular signals at different stages of development and in response to changing environmental conditions. Previously protein turnover studies employing metabolic labeling approaches with stable isotopes were performed with primarily manual data collection and processing. We developed a method that used ~(15)N and ~(13)C as general metabolic labels, coupled with high-throughput mass spectrometry, for the measurement of dynamic protein turnover in plants. We now report important improvements of this method that allows rapid and accurate protein turnover measurements using an isotopic labeling followed by dilution approach as well as an incorporation approach.