Supplementary Materials

The PDF file includes:

  • Fig. S1. RAS isoform-specific HVRs.
  • Fig. S2. There is no major interaction between unfarnesylated KRAS4b (1–185) and CaM, independent of bound nucleotide.
  • Fig. S3. A new class of noncanonical CaM binding targets: Singly lipidated polybasic peptides.
  • Fig. S4. Chemical shift analysis of CaM residues upon farnesol binding.
  • Fig. S5. Farnesol and CaM do not interact in the absence of Ca2+.
  • Fig. S6. Farnesyl compound titrations into CaM.
  • Fig. S7. KRAS4b-FMe and farnesol perturbations of 15N-CaM share a characteristic pattern.
  • Fig. S8. Structure comparison of CaM conformations bound to FCME and CAP23/NAP22 polybasic myristoylated peptide.
  • Fig. S9. Lipid orientation in CaM:FCME complex structure.
  • Fig. S10. Solution NMR PRE validation of CaM-FCME crystal structure.
  • Fig. S11. Membrane-based PRE probe for KRAS4b-FMe interaction with CaM in solution.
  • Fig. S12. KRAS4b retains function in the CaMeRAS chimera.
  • Fig. S13. Calcium-dependent change in emission spectra of CaMeRAS chimera FRET protein.
  • Fig. S14. Tracking cellular localization of CaMeRASWT in HeLa cells after ionomycin treatment.
  • Fig. S15. Cellular image segmentation for FRET histogram analysis by Gaussian modeling.
  • Fig. S16. Fluorescence lifetime of mTq2 of CaMeRASWT is reduced by histamine stimulation in HeLa cells.
  • Fig. S17. ATP-evoked Ca2+ signal induces CaMeRASWT internalization and FRET change in HEK293 cells.
  • Fig. S18. The nucleotide and amino acid sequences of CaMeRASWT chimeric FRET construct.
  • Table S1. Peak values of FRET histograms and the numbers of the ROIs analyzed.
  • Table S2. P values from Student’s t tests comparing Gaussian curve models of FRET histogram data.
  • Table S3. Primer sequences used for plasmid construction.
  • Legends for movies S1 to S5
  • Legends for data files S1 to S6

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.avi format). Co-imaging CaMeRASWT and calcium response to histamine stimulation in HeLa cells.
  • Movie S2 (.avi format). Slow motion highlight: Co-imaging CaMeRASWT and calcium response to histamine stimulation in HeLa cells.
  • Movie S3 (.avi format). Coimaging CaMeRASWT and calcium response to histamine stimulation in HeLa cells and calcium chelation.
  • Movie S4 (.avi format). Coimaging CaMeRAS185A and calcium response to histamine stimulation in HeLa cells.
  • Movie S5 (.avi format). Coimaging CaMeRAS1234Q and calcium response to histamine stimulation in HeLa cells.
  • Data file S1 (Microsoft Excel format). CaM-farnesol binding CSP by residue.
  • Data file S2 (Microsoft Excel format). CaM-FCME PRE.
  • Data file S3 (Microsoft Excel format). KRAS4b-ND PRE.
  • Data file S4 (Microsoft Excel format). In vitro CaMeRAS Ca2+ titration.
  • Data file S5 (Microsoft Excel format). Relative fluorophore intensity in vitro as a function of Ca2+ titration for CaMeRAS.
  • Data file S6 (.zip file). In vivo FRET from CaMeRAS contructs in response to histamine stimulation and Ca2+ influx.