Hybridization of Glyceraldehyde-3-phosphate Dehydrogenase in Borate
1976 年 79 巻 6 号 p. 1287-1295
詳細
1976 年 79 巻 6 号 p. 1287-1295
Conditions for the hybridization of glyceraldehyde-3-phosphate dehydrogenase (GPD) [EC 1.2.1.12] in the presence of dilute borate were examined with horseshoe crab and rabbit GPDs. Hybridization was strongly dependent upon pH, borate concen-tration, and temperature. The optimum medium for hybridization was 10mM Tris-HCl-1mM 2-mercaptoethanol-1mM EDTA containing 10-20mM borate, pH 8.5-9.0. Hybridization was performed by incubation of two
electrophoretically distinct GDPs at 30° for 3-15 hr in the above medium at a protein concentration of 1-2mg/ml.
The time course of hybridization was analyzed under the optimized conditions. Symmetrical A2B2-type hybrid appeared only 5min after incubation for 1 hr.
Hybridization of GPDs from 7 different species was examined under the optimal conditions. Hybridization was detected with rabbit-horseshoe crab, yeast-rabbit, yeast-chicken, and chicken-horseshoe crab combinations.
Subunit-subunit interaction, the mechanism of hybridization, and the structure of GPD are discussed based on the results obtained.
