Bioinformatics Analysis of Top-Down Mass Spectrometry Data with ProSight Lite.
Traditional bottom-up mass spectrometry-based proteomics relies on the use of an enzyme, often trypsin, to generate small peptides (typically < 25 amino acids long). In top-down proteomics, proteins remain intact and are directly measured within the mass spectrometer. This technique, while inherently simpler than bottom-up proteomics, generates data which must be processed and analyzed using software tools "purpose-built" for the job. In this chapter, we will show the analysis of intact protein spectra through deconvolution, deisotoping, and searching with ProSight Lite, a free, vendor-agnostic tool for the analysis of top-down mass spectrometry data. We will illustrate with two examples of intact protein fragmentation spectra and discuss the iterative use of the software to characterize proteoforms and discover the sites of post-translational modifications.
['DeHart|Caroline J|CJ|', 'Fellers|Ryan T|RT|', 'Fornelli|Luca|L|', 'Kelleher|Neil L|NL|', 'Thomas|Paul M|PM|']
D000818:Animals D019295:Computational Biology D057225:Data Mining D006801:Humans D013058:Mass Spectrometry D011506:Proteins D040901:Proteomics D012984:Software D014584:User-Computer Interface D064878:Web Browser
Bioinformatics Top-down Mass spectrometry ProSight Lite Proteomics Intact protein