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Updated: Feb 27, 2026
Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks
Published on: December 24, 2016
Analysis of K-Ras Interactions by Biotin Ligase Tagging
Christopher Ritchie1, Andrew Mack1, Logan Harper1
1Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, U.S.A.
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 proximity-dependent 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.
