Physical estimation of triplet phases from two new proteins

Three-beam interference experiments have been performed with crystals of two glycosidases: guinea-fowl hexagonal lysozyme, MW 14.3 kDa, and C. thermocellum endoglucanase CelA, MW 40 kDa. In both cases triplet phases could be estimated. Experimental parameters and details of the procedure are presented along with some examples of the results. The average differences between the estimated phases and those calculated from the crystallographic refinements were 17.9 and 15.9°, respectively. A brief discussion of alternative methods for physical phase acquisition is given, including possible strategies for the measurement and application of experimental phases in macromolecular crystallography.