@inbook{10ff20d1e9e14522a5b9f8fe3b35ee2d,
title = "Precise Characterization of KRAS4B Proteoforms by Combining Immunoprecipitation with Top-Down Mass Spectrometry",
abstract = "The characterization of biologically relevant post-translational modifications (PTMs) on KRAS4B has historically been carried out through methodologies such as immunoblotting with PTM-specific antibodies or peptide-based proteomic methods. While these methods have the potential to identify a given PTM on KRAS4B, they are incapable of characterizing or distinguishing the different molecular forms or proteoforms of KRAS4B from those of related RAS isoforms. We present a method that combines immunoprecipitation of KRAS4B with top-down mass spectrometry (IP-TDMS), thus enabling the precise characterization of intact KRAS4B proteoforms. We provide detailed protocols for the IP, LC-MS/MS, and data analysis comprising a successful IP-TDMS assay in the contexts of cancer cell lines and tissue samples.",
keywords = "Immunoprecipitation, Intact protein, Isoform, Proteoform, RAS, Top-down mass spectrometry",
author = "Adams, {Lauren M.} and DeHart, {Caroline J.} and Kelleher, {Neil L.}",
note = "Funding Information: We thank Kevin Haigis for providing an agarose bead-based IP protocol and the following coauthors of the Ntai et al. [7] manuscript for their significant contributions to the initial IP-TDMS method development: Ioanna Ntai and Luca Fornelli. This work was supported by federal funds from the National Cancer Institute (Office of Cancer Clinical Proteomics Research), National Institutes of Health, under Contract HHSN261200800001E, and Lei-dos Biomedical Research under Contract HHSN261200800001E and was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under National Institutes of Health Grant P41 GM108569. L.M.A. is supported by T32GM008382. Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2021",
doi = "10.1007/978-1-0716-1190-6_3",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "47--64",
booktitle = "Methods in Molecular Biology",
}
