Calmodulin disrupts plasma membrane localization of farnesylated KRAS4b by sequestering its lipid moiety.

KRAS4b is a small guanosine triphosphatase (GTPase) protein that regulates several signal transduction pathways that underlie cell proliferation, differentiation, and survival. KRAS4b function requires prenylation of its C terminus and recruitment to the plasma membrane, where KRAS4b activates effector proteins including the RAF family of kinases. The Ca ²⁺ -sensing protein calmodulin (CaM) has been suggested to regulate the localization of KRAS4b through direct, Ca ²⁺ -dependent interaction, but how CaM and KRAS4b functionally interact is controversial. Here, we determined a crystal structure, which was supported by solution nuclear magnetic resonance (NMR), that revealed the sequestration of the prenyl moiety of KRAS4b in the hydrophobic pocket of the C-terminal lobe of Ca ²⁺ -bound CaM. Our engineered fluorescence resonance energy transfer (FRET)-based biosensor probes (CaMeRAS) showed that, upon stimulation of Ca ²⁺ influx by extracellular ligands, KRAS4b reversibly translocated in a Ca ²⁺ -CaM-dependent manner from the plasma membrane to the cytoplasm in live HeLa and HEK293 cells. These results reveal a mechanism underlying the inhibition of KRAS4b activity by Ca ²⁺ signaling pathways.
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