Science Advances

Supplementary Materials

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  • fig. S1. 1H-13C methyl-TROSY spectra of apo, nucleotide-bound (ATPγC), and ternary (ATPγN/PKI5-24 and ATPγN/PLN1–19) forms of 2H,13C-isoleucine, leucine, and valine (IVL)–labeled PKA-C.
  • fig. S2. Plots of the IVL methyl group CSPs upon ligand binding.
  • fig. S3. Mapping of the CSPs of the IVL methyl side-chain groups onto the crystal structure of PKA-C (PDB: 1ATP).
  • fig. S4. Statistical analysis of the chemical shift changes.
  • fig. S5. CHESCA (39) correlation matrix showing the degree of correlated chemical shift changes for the methyl side chains of IVL residues.
  • fig. S6. Global conformational transitions mapped via linear CSPs probed by amide backbone resonances and methyl group resonances.
  • fig. S7. Fast time scale (picosecond to nanosecond) conformational dynamics of the kinase upon ligand binding.
  • fig. S8. Thermodynamics of ligand binding for PKA-C as measured by isothermal titration calorimetry (ITC).
  • fig. S9. Slow time scale (microsecond to millisecond) conformational dynamics of the kinase.
  • fig. S10. Slow time scale conformational dynamics of the kinase.
  • fig. S11. Synchronous dynamics occurring in the highly conserved hydrophobic core.
  • fig. S12. Location of I150 at the interface between different communities of PKA-C.
  • fig. S13. Bridging residues connect the R-spine and C-spine at the PKA hydrophobic core.
  • fig. S14. Western blot–based activity assay.
  • fig. S15. Assembly of the R-spine for active protein kinases.
  • fig. S16. Expression and purification of recombinant 2H, 15N, 13CH3-ILV, and PKA-C from E. coli bacteria.
  • table S1. Classification of residues undergoing correlated chemical shift changes and their respective location in a specific community as identified by community map analysis.
  • table S2. The dynamic light scattering data for three different forms of PKA-C.
  • table S3. T2 and S2 values for methyl side-chain groups of apo PKA-C.
  • table S4. T2 and S2 values for methyl side-chain groups of the ATPγC-bound state of PKA-C.
  • table S5. T2 and S2 values for methyl side-chain groups of the ATPγN/PKI5-24-bound state of PKA-C.
  • table S6. Group fits of CPMG dispersion curves measured at 700 and 850 MHz of the apo form of PKA-C.
  • table S7. Group fits of the CPMG relaxation dispersion curves measured at 700 and 850 MHz of the ATPγC form of PKA-C.
  • table S8. Single-quantum individual site fits of CPMG relaxation dispersion curves measured at 700 and 850 MHz of the apo form of PKA-C.
  • table S9. Individual fits of the CPMG dispersion curves measured at 700 and 50 MHz of the ATPγC-bound state of PKA-C.
  • table S10. Single-quantum individual site fits of CPMG curves at 700 and 850 MHz of the ATPγN/PKI5-24-bound state of PKA-C.
  • table S11. Approximate Rex values from two points of the CPMG experiment for the ATPγN/PLN1–19-bound form of PKA-C.
  • Reference (54)

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