Comparison of the independent solvent structures of dimeric alpha-chymotrypsin with themselves and with gamma-chymotrypsin.

The solvent structure of alpha-chymotrypsin has been determined in the restrained least squares refinement (1.67-A resolution) of the dimeric molecule (Blevins, R. A., and Tulinsky, A. (1985) J. Biol. Chem. 260, 4264-4275). A total of 247 water molecules reduced the R-factor by 0.039 to 0.179. The average occupancy of solvent is 0.77 and the average isotropic thermal parameter is 22 A2. About 80% of the solvent is around the surface, 10% is in the dimer interface, and 10% is interior. There are 49 pairs of water molecules related by 2-fold noncrystallographic symmetry (within 1.0 A) and 199 waters that can potentially hydrogen bond with protein or themselves. The specificity sites contain 5 water molecules, 2 of which are displaced by substrate binding. The remainder probably aid in identifying and positioning the latter for catalysis. Four of these waters also occur in gamma-chymotrypsin. Considering the water structure in the dimer interface region of alpha-chymotrypsin with that of gamma-chymotrypsin reveals that about two-thirds of the solvent in this region is lost on dimerization. Last, 4 of the water molecules of alpha-chymotrypsin have been identified to be sulfate ions from a difference map based on crystals with selenate exchanged mother liquor.