Fast Proteome Identification and Quantification from Data-Dependent Acquisition - Tandem Mass Spectrometry using Free Software Tools

Abstract: Identification of nearly all proteins in a system using data-dependent acquisition (DDA) mass spectrometry has become routine for simple organisms, such as bacteria and yeast. Still, quantification of the identified proteins may be a complex process and require multiple different software packages. This protocol describes identification and label-free quantification of proteins from bottom-up proteomics experiments. This method can be used to quantify all the detectable proteins in any DDA dataset collected with high-resolution precursor scans. This protocol may be used to quantify proteome remodeling in response to a drug treatment or a gene knockout. Notably, the method uses the latest and fastest freely-available software, and the entire protocol can be completed in a few hours with data from organisms with relatively small genomes, such as yeast or bacteria.