Molecular organization and structural stability of beta s-crystallin from calf lens

beta s-Crystallin has been purified to homogeneity. Its structural features and conformational behavior have been studied in solution. Protein secondary structure was estimated by curve fitting of far-UV circular dichroism spectra, which gave 16% alpha-helix, 45% beta-sheet, 12% bends, and 27% remainders. This result indicates that the structural organization of beta s-crystallin is reasonably similar to that of other beta and gamma family members. A comparison assessed between beta s- and gamma 2-crystallin by the use of predictive methods (flexibility and volume plots) reveals that the two proteins differ in respect to their local flexibility and packing, although they show similar overall organization. The interdomain and the C-terminal regions were found to be more flexible in beta s-crystallin. This finding can be explained by the presence of smaller amino acid residues within these structural districts. The location of one out of four tryptophans, i.e., Trp-162, in a flexible and exposed region of the protein was found to be the origin of the fluorescence heterogeneity. In fact, the fluorescence emission maximum of the native protein, centered at 328 nm, is due to two emitting centers, whose emission maxima are located at 323 and 330 nm, respectively, as evidenced by acrylamide quenching of fluorescence. The effect of perturbing agents, such as pH and guanidine hydrochloride, on the conformational behavior of beta s has also been evaluated by numerous spectroscopic techniques. The range of pH stability was between 6.5 and 8.(ABSTRACT TRUNCATED AT 250 WORDS)