Browsing by Author "DeHart, Caroline J"

Filter results by typing the first few letters
Now showing 1 - 2 of 2
  • No Thumbnail Available
    Item
    Bioinformatics Analysis of Top-Down Mass Spectrometry Data
    (Methods in Molecular Biology, Springer) DeHart, Caroline J; Fellers, Ryan T; Fornelli, Luca; Kelleher, Neil L; Thomas, Paul M
    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 proteins from deconvolution and deisotoping through analysis 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 spectra and discuss the iterative use of the software to characterize proteoforms and to discover the sites of post-translational modifications.
  • No Thumbnail Available
    Item
    Identification and characterization of human proteoforms by top-down LC-21 tesla FT-ICR mass spectrometry
    (American Chemical Society, 2016) Anderson, Lissa C; DeHart, Caroline J; Kaiser, Nathan K; Fellers, Ryan T; Smith, Donald F; Greer, Joseph B; LeDuc, Richard D; Blackney, Gregory T; Thomas, Paul M; Kelleher, Neil L; Hendrickson, Christopher L
    The National High Magnetic Field Laboratory (NHMFL) recently installed a 21T FT-ICR mass spectrometer, which is part of the NHMFL FT-ICR User Facility and available to all qualified users. Here, we demonstrate top-down LC-21 T FT-ICR MS/MS of intact proteins derived from human colorectal cancer cell lysate. We identify a combined total of 684 unique protein entries observed as 3,238 unique proteoforms at a 1% false discovery rate, based on rapid, data-dependent acquisition of collision-induced and electron-transfer dissociation tandem mass spectra from 40 LC-MS/MS experiments.