Expression of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in escherichia coli

Lin Li, Wei Zhe Yao, Alex J Lange, Simon J. Pilkis, Meng Qiu Dong, Yong Yin, Gen Jun Xu

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase was expressed in E. coli by using a pET3a T7 RNA polymerase-based expression system and was purified to homogeneity. The kinase and bisphosphatase of the expressed bifunctional enzyme had kinetic properties identical to those of the native chicken liver enzyme. However, the kinase activity of the chicken liver enzyme was 7-fold higher, while the bisphosphatase activity was 50 percent lower than those of the rat liver enzyme. Cys-256 of the rat liver bisphosphatase domain is not conserved in the chicken liver enzyme. A site-directed mutation was engineered at Cys-256 of the rat liver enzyme and the results indicate that the variation of this residue is not responsible for the difference in fructose-2,6-bisphosphatase activity between the rat and chicken liver enzymes. It is postulated that the difference in the kinase/bisphosphatase activity ratios of these two enzymes results from differences in their NH2 -terminal regions.

Original languageEnglish (US)
Pages (from-to)883-893
Number of pages11
JournalBiochemical and Biophysical Research Communications
Volume209
Issue number3
DOIs
StatePublished - Apr 26 1995

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