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Human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase: its isozymic form, expression and
R Sakakibara1, M Uemura, T Hirata
1Department of Biochemistry, School of Pharmaceutical Sciences, Nagasaki University, Japan.
Abstract:
The nucleotide sequence of 1981 bp cDNA containing the entire coding region of a human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase was determined. The sequence encodes 469 amino acids and, based on homology to the rat testis enzyme, appears to be the testis-type isozyme expressed in placenta. The enzyme was expressed in Escherichia coli BL21 (DE3) by using a T7 RNA polymerase-based expression system and purified to homogeneity. The expressed enzyme was bifunctional with specific activities of 75 and 80 mU/mg of kinase and phosphatase, respectively. Kinetic parameters of the expressed enzyme are similar to those of the rat testis enzyme.