Coreapplication of tmtpro™ 16plex mass tagging strategy to quantitatively characterize heat-stable proteome of escherichia coli.
The Mass Spectrometry Core Facility at the Biotechnology Center has been working closely with Dr. Silvia Cavagnero’s lab in the Department of Chemistry to unravel the dynamics of biomolecules in their native environment. In this particular experimental approach, we have taken advantage of the ThermoFisher Scientific TMTPro 16plex peptide-tagging reagents to analyze 16 individual samples in a single multiplexed, high-capacity nanoLC-MS/MS run. Several thousands of peptide reads were generated and abundances from 16 pre-mixed samples were assigned. We have been able to unequivocally identify and quantify 1,875 protein groups and further evaluate individual protein thermal stabilities upon heat treatment. Our ability to label proteomes at the early stages of sample preparation allowed for immediate mixing of multiple treatments and biological replicates leading to superior technical reproducibility at significant overall cost reduction associated with streamlined sample processing, sample multiplexing, and decreased instrument time.