Ras membrane orientation and nanodomain localization generate isoform diversity

Daniel Abankwa, AA Gorfe, K Inder, JF Hancock

Research output: Contribution to journalArticleScientificpeer-review

169 Citations (Scopus)


The structural elements encoding functional diversity among Ras GTPases are poorly defined. The orientation of the G domain of H-ras with respect to the plane of the plasma membrane is recognized by the Ras binding domain of C-Raf, coupling orientation to MAPK activation. We now show that two other proteins, phosphoinositide-3-kinase-alpha and the structurally unrelated galectin-1, also recognize G-domain orientation. These results rationalize the role of galectin-1 in generating active GTP-H-ras signaling nanoclusters. However, molecular dynamics simulations of K-ras membrane insertion and fluorescence lifetime imaging microscopy (FLIM)-Forster resonance energy transfer (FRET) imaging of the effector interactions of N-Ras, K-Ras, and M-ras suggest that there are two hyperactive, signaling-competent orientations of the Ras G domain. Mutational and functional analyses establish a clear relationship between effector binding and the amphilicities of helix alpha 4 and the C-terminal hypervariable region, thus confirming that these structural elements critically tune the orientation of the Ras G domain. Finally, we show that G-domain orientation and nanoclustering synergize to generate Ras isoform specificity with respect to effector interactions.
Original languageUndefined/Unknown
Pages (from-to)1130–1135
Number of pages6
JournalProceedings of the National Academy of Sciences
Issue number3
Publication statusPublished - 2010
MoE publication typeA1 Journal article-refereed


  • FRET
  • nanocluster
  • signal transduction
  • small G protein

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