Crystallization of chicken egg white lysozyme from assorted sulfate salts

Chicken egg white lysozyme has been found to crystallize from ammonium, sodium, potassium, rubidium, magnesium, and manganese sulfates at acidic and basic pH, with protein concentrations from 60 to 190 mg/ml. Crystals have also been grown at 4 degrees C in the absence of any other added salts using isoionic lysozyme which was titrated to pH 4.6 with dilute sulfuric acid. Four different crystal forms have been obtained, depending upon the temperature, protein concentration, and precipitating salt employed. Crystals grown at 15 degrees C were generally tetragonal, with space group P4(3)2(1)2. Crystallization at 20 degrees C typically resulted in the formation of orthorhombic crystals, space group P2(1)2(1)2(1). The tetragonal <-> orthorhombic transition appeared to be a function of both the temperature and protein concentration, occurring between 15 and 20 degrees C and between 100 and 125 mg/ml protein concentration. Crystallization from 1.2 M magnesium sulfate at pH 7.8 gave a trigonal crystal, space group P3(1)2(1), a = b = 87.4, c = 73.7, gamma = 120 degrees, which diffracted to 2.8 Angstrom. Crystallization from ammonium sulfate at pH 4.6, generally at lower temperatures, was also found to result in a monoclinic form, space group C2, a = 65.6, b = 95.0, c = 41.2, beta = 119.2 degrees. A crystal of similar to 0.2 x 0.2 x 0.5 mm grown from bulk solution diffracted to similar to 3.5 Angstrom. (C) 1999 Elsevier Science B.V. All rights reserved.