@article {RITCHIE225, author = {CHRISTOPHER RITCHIE and ANDREW MACK and LOGAN HARPER and AYNA ALFADHLI and PHILIP J.S. STORK and XIAOLIN NAN and ERIC BARKLIS}, title = {Analysis of K-Ras Interactions by Biotin Ligase Tagging}, volume = {14}, number = {4}, pages = {225--239}, year = {2017}, publisher = {International Institute of Anticancer Research}, 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 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.}, issn = {1109-6535}, URL = {https://cgp.iiarjournals.org/content/14/4/225}, eprint = {https://cgp.iiarjournals.org/content/14/4/225.full.pdf}, journal = {Cancer Genomics \& Proteomics} }