期刊
NATURE METHODS
卷 11, 期 5, 页码 585-592出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.2895
关键词
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资金
- Ontario Genomics Institute (303547)
- Canadian Institutes of Health Research [Catalyst-NHG94491, PPP-125785]
- Canadian Foundation for Innovation (IOF-LOF)
- Natural Sciences and Engineering Research Council of Canada (RGPIN) [372393-12]
- Canadian Cystic Fibrosis Foundation [300348]
- Canadian Cancer Society [2010-700406]
- Novartis and University Health Network [GL2-01-018]
- Fonds zur Forderung der wissenschaftlichen Forschung-Erwin Schrodinger postdoctoral fellowship
Cell signaling, one of key processes in both normal cellular function and disease, is coordinated by numerous interactions between membrane proteins that change in response to stimuli. We present a split ubiquitin based method for detection of integral membrane protein-protein interactions (PPIs) in human cells, termed mammalian-membrane two-hybrid assay (MaMTH). We show that this technology detects stimulus (hormone or agonist)-dependent and phosphorylation-dependent PPIs. MaMTH can detect changes in PPIs conferred by mutations such as those in oncogenic ErbB receptor variants or by treatment with drugs such as the tyrosine kinase inhibitor erlotinib. Using MaMTH as a screening assay, we identified CRKII as an interactor of oncogenic EGFR(L858R) and showed that CRKII promotes persistent activation of aberrant signaling in non small cell lung cancer cells. MaMTH is a powerful tool for investigating the dynamic interactomes of human integral membrane proteins.
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