The molecular mechanics Poisson Boltzmann and surface area method (MMPBSA) was used to estimate the binding affinity between segments of cTn with the rest of the protein complex. MMPBSA utilises molecular mechanics, implicit continuum solvent model, and solvent accessible surface area to estimate the enthalpy of binding . The atomic radii set for the Poisson Boltzmann calculation was chosen to be mbondi2. MMPBSA.py , part of AmberTools, was used to carry out the calculations. No entropic contribution was calculated.