Human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase: Its isozymic form, expression and characterization
Human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase: Its isozymic form, expression and characterization
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DOI：
10.1271/bbb.61.1949
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发表时间：
1997-11-01
影响因子：
1.6
通讯作者：
Kato, M
中科院分区：
文献类型：
--
作者：
Sakakibara, R;Uemura, M;Kato, M
测定了人胎盘果糖-6-磷酸，2-激酶/2，6-二磷酸果糖-2，6-二磷酸酶编码区全长的1981个碱基的核苷酸序列。该序列编码469个氨基酸，根据与大鼠睾丸酶的同源性，似乎是在胎盘中表达的睾丸型同工酶。利用基于T7RNA聚合酶的表达系统在大肠杆菌BL21(DE3)中表达了该酶，并进行了纯化。表达的酶具有双功能，比活力分别为75和80mU/mg的激酶和磷酸酶。表达的酶的动力学参数与大鼠睾丸酶相似。
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 80mU/mg of kinase and phosphatase, respectively. Kinetic parameters of the expressed enzyme are similar to those of the rat testis enzyme.