Two Distinct Structures of Membrane-Associated Homodimers of GTP- and GDP-Bound KRAS4B Revealed by Paramagnetic Relaxation Enhancement

Angew Chem Int Ed Engl. 2020 Jun 26;59(27):11037-11045. doi: 10.1002/anie.202001758. Epub 2020 Apr 30.


KRAS homo-dimerization has been implicated in the activation of RAF kinases, however, the mechanism and structural basis remain elusive. We developed a system to study KRAS dimerization on nanodiscs using paramagnetic relaxation enhancement (PRE) NMR spectroscopy, and determined distinct structures of membrane-anchored KRAS dimers in the active GTP- and inactive GDP-loaded states. Both dimerize through an α4-α5 interface, but the relative orientation of the protomers and their contacts differ substantially. Dimerization of KRAS-GTP, stabilized by electrostatic interactions between R135 and E168, favors an orientation on the membrane that promotes accessibility of the effector-binding site. Remarkably, "cross"-dimerization between GTP- and GDP-bound KRAS molecules is unfavorable. These models provide a platform to elucidate the structural basis of RAF activation by RAS and to develop inhibitors that can disrupt the KRAS dimerization. The methodology is applicable to many other farnesylated small GTPases.

Keywords: KRAS; NMR spectroscopy; dimerization; membrane proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Dimerization
  • Guanosine Diphosphate / metabolism*
  • Guanosine Triphosphate / metabolism*
  • Humans
  • Magnetic Resonance Spectroscopy / methods
  • Molecular Dynamics Simulation
  • Proto-Oncogene Proteins p21(ras) / chemistry
  • Proto-Oncogene Proteins p21(ras) / metabolism*


  • KRAS protein, human
  • Guanosine Diphosphate
  • Guanosine Triphosphate
  • Proto-Oncogene Proteins p21(ras)