Investigation of the solution structure of chymotrypsin inhibitor 2 using molecular dynamics: Comparison to X-ray crystallographic and NMR data

The native solution structure and dynamics of chymotrypsin inhibitor 2 (CI2) have been studied using a long (5.3 ns) molecular dynamics (MD) simulation without any imposed restraints, The majority of the experimentally observed spin-spin coupling constants, short- and long-range nuclear Overhauser effect (NOE) cross peaks and the amide hydrogen exchange behavior were reproduced by the MD simulation. This good correspondence suggests that the major structural features of the protein during the simulation are representative of the true protein structure in solution. Two water molecules formed hydrogen bond bridges between beta(2) and beta(3), in agreement with X-ray crystallographic data and a recent reassessment of the solution structure using time-averaged NMR restraints during MD refinement. The active-site loop of the protein displayed the greatest structural changes and the highest mobility. When this loop region was excluded, the average C-alpha r.m,s, deviation of the simulated solution structures from the crystal structure was similar to 1.5 Angstrom from 0.5 to 5.3 ns, There is structural heterogeneity in particular regions of the NMR-derived solution structures, ,which could be a result of imprecision or true internal motion, A study of the distribution of mobility through the protein allows us to distinguish between these two alternatives. In particular, deviations in the active-site loop appear to be a result of heightened mobility, which is also supported by good correspondence between calculated and experimental S-2 N-H order parameters. On the other hand, other ill-defined regions of the NMR-derived structures are well defined in the simulation and are probably the result of a lack of structural restraints (i.e, NOEs), as opposed to reflecting the true mobility.