Abstract
Background: Mutations of the human K-Ras 4B (K-Ras) G protein are associated with a significant proportion of all human cancers. Despite this fact, a comprehensive analysis of K-Ras interactions is lacking. Our investigations focus on characterization of the K-Ras interaction network. Materials and Methods: We employed a biotin ligase-tagging approach, in which tagged K-Ras proteins biotinylate neighbor proteins in a proximitydependent fashion, and proteins are identified via mass spectrometry (MS) sequencing. Results: In transfected cells, a total of 748 biotinylated proteins were identified from cells expressing biotin ligase-tagged K-Ras variants. Significant differences were observed between membrane-associated variants and a farnesylation-defective mutant. In pancreatic cancer cells, 56 K-Ras interaction partners were identified. Most of these were cytoskeletal or plasma membrane proteins, and many have been identified previously as potential cancer biomarkers. Conclusion: Biotin ligase tagging offers a rapid and convenient approach to the characterization of K-Ras interaction networks.
Original language | English (US) |
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Pages (from-to) | 225-239 |
Number of pages | 15 |
Journal | Cancer Genomics and Proteomics |
Volume | 14 |
Issue number | 4 |
DOIs | |
State | Published - Jul 1 2017 |
Keywords
- Pancreatic cancer
- Proximity mapping
- Subcellular localization
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Genetics
- Cancer Research