Histidine residues in alpha-crystallin are not all available for chemical modification and acid-base titration.

We have determined the number of histidine residues available for chemical modification with the specific reagent diethylpyrocarbonate in both bovine and goat alpha-crystallins. Results indicate that there are two distinctly different classes of histidine residues in the native protein. Out of 300 total histidine residues in the protein (on the basis of 800-kDa protein molecular weight) about 170 +/- 2 residues have been found to be modified by the reagent. The remaining 130 +/- 2 residues are modified when the protein is partially denatured in 1.5 M guanidine hydrochloride. The H(+)-titration behavior of the histidine residues in the protein corroborates this result. The observed differential accessibility of histidine residues may be important in maintaining the surface hydrophobicity of the aggregate as well as in stabilizing its quaternary structure.