Molecular cloning, sequence analysis, and expression of a human liver cDNA coding for fructose-6-P,2-kinase:Fructose-2,6-bisphosphatase

Joseph Algaier, Kosaku Uyeda

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

A cDNA coding for 378 amino acids from the C-terminus of the human liver bifunctional enzyme, Fructose-6-phosphate,2-kinase:Fructose-2,6-bisphosphatase was isolated, sequenced, and expressed in E. coli K38. The expressed protein, identified by specific immunoassay, showed Fru 2,6-bisphosphatase activity but no Fru 6-P,2-kinase activity, demonstrating directly that the Fru 2,6-bisphosphatase activity resides in the C-terminal region. The Km for Fru 2,6-P2 was 4.3 μM. Fru 6-P was a noncompetitive inhibitor (Ki = 2.9 μM), and formed a phosphorylated intermediate when incubated with Fru 2,6[2-32P]P2. The subunit Mr of the enzyme was 36,600, and the active enzyme showed Mr=37,000 by gel filtration.

Original languageEnglish (US)
Pages (from-to)328-333
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume153
Issue number1
DOIs
StatePublished - May 31 1988

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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