Mapping the KRAS proteoform landscape in colorectal cancer identifies truncated KRAS4B that decreases MAPK signaling

Lauren M. Adams, Caroline Jane DeHart, Bryon S. Drown, Lissa C. Anderson, William Bocik, Emily S. Boja, Tara M. Hiltke, Christopher L. Hendrickson, Henry Rodriguez, Michael Caldwell, Reza Vafabakhsh, Neil L Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

The KRAS gene is one of the most frequently mutated oncogenes in human cancer and gives rise to two isoforms, KRAS4A and KRAS4B. KRAS post-translational modifications (PTMs) have the potential to influence downstream signaling. However, the relationship between KRAS PTMs and oncogenic mutations remains unclear, and the extent of isoform-specific modification is unknown. Here, we present the first top–down proteomics study evaluating both KRAS4A and KRAS4B, resulting in 39 completely characterized proteoforms across colorectal cancer cell lines and primary tumor samples. We determined which KRAS PTMs are present, along with their relative abundance, and that proteoforms of KRAS4A versus KRAS4B are differentially modified. Moreover, we identified a subset of KRAS4B proteoforms lacking the C185 residue and associated C-terminal PTMs. By confocal microscopy, we confirmed that this truncated GFP-KRAS4BC185∗ proteoform is unable to associate with the plasma membrane, resulting in a decrease in mitogen-activated protein kinase signaling pathway activation. Collectively, our study provides a reference set of functionally distinct KRAS proteoforms and the colorectal cancer contexts in which they are present.

Original languageEnglish (US)
Article number102768
JournalJournal of Biological Chemistry
Volume299
Issue number1
DOIs
StatePublished - Jan 2023

Keywords

  • KRAS
  • MAPK pathway
  • RAS protein
  • colorectal cancer
  • immunoprecipitation
  • post-translational modification
  • proteoform
  • top–down proteomics

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry
  • Cell Biology

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