Fructose-bisphosphatase, Zinc-Free, from Rabbit Liver

Margaret M. Demaine, Carol A. Caperelli, Stephen J. Benkovic

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


This chapter describes an assay method and the purification procedure for fructose-bisphosphatase (FBPase) from rabbit liver. The enzyme is assayed spectrophotometrically by following the rate of NADPH accumulation at 340 nm in the presence of excess glucose-6-phosphate dehydrogenase and glucose-6-phosphate isomerase. Neutral FBPase is purified from frozen livers of young, 24-hr-fasted rabbits. For Zn analysis, atomic absorption spectroscopy is employed to measure the concentration of the stock Zn2+ solution and the levels of Zn2+ (and Mn2+) in buffer solutions, water, and FBPase samples. Removal of Zn2+ from FBPase involves various steps; the removal of the tightly bound Zn2+ does not irreversibly alter the FBPase structure. However, experimental studies of other properties of zinc-free FBPase are technically difficult, because the exposure of the enzyme to buffer or other reagents immediately results in binding of the adventitious Zn2+ by FBPase.

Original languageEnglish (US)
Pages (from-to)327-329
Number of pages3
JournalMethods in enzymology
Issue numberC
StatePublished - Jan 1 1982

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology


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