CALCULATION OF THE RELATIVE BINDING FREE-ENERGY OF 2'GMP AND 2'AMP TO RIBONUCLEASE-T1 USING MOLECULAR-DYNAMICS FREE-ENERGY PERTURBATION APPROACHES

We present a calculation of the relative changes in binding free energy between the complex of ribonuclease T1 (RNase T1) with its inhibitor 2′-guanosine monophosphate (2′GMP) and that of RNase T1-2′-adenosine monophosphate (2′AMP) by means of a thermodynamic perturbation method implemented with molecular dynamics. Using the available crystal structure of the RNase T1-2′GMP complex, the structure of the RNase T1-2′AMP complex was obtained as a final structure of the perturbation calculation. The calculated difference in the free energy of binding (ΔΔGbind) was 2.76 kcal/mol. This compares well with the experimental value of 3.07 kcal/mol. The encouraging agreement in ΔΔGbind suggests that the interactions of inhibitors with the enzyme are reasonably represented. Energy component analyses of the two complexes reveal that the active site of RNase T1 electrostatically stabilizes the binding of 2′GMP more than that of 2′AMP by 44 kcal/mol, while the van der Waals' interactions are similar in the two complexes. The analyses suggest that the mutation from Glu46 to Gln may lead to a preference of RNase T1 for adenine in contrast to the guanine preference of the wild-type enzyme. Although the molecular dynamics equilibration moves the atoms of the RNase T1-2′GMP system about 0.9 Å from their X-ray positions and the mutation of the G to A in the active site increases the deviation from the X-ray structure, the mutation of the A back to G reduces the deviation. This and the agreement found for ΔΔGbind suggest that the molecular dynamics/free energy perturbation method will be useful for both energetic and structural analysis of protein-ligand interactions. © 1990.