Fructose-bisphosphatase, Zinc-Free, from Rabbit Liver

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

Research output: Contribution to journalArticle

Abstract

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
Volume90
Issue numberC
DOIs
StatePublished - Jan 1 1982

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Fructose-Bisphosphatase
Liver
Zinc
Rabbits
Buffers
Glucose-6-Phosphate Isomerase
Atomic spectroscopy
Glucosephosphate Dehydrogenase
Enzymes
NADP
Absorption spectroscopy
Purification
Assays
Spectrum Analysis
Water

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Demaine, Margaret M. ; Caperelli, Carol A. ; Benkovic, Stephen. / Fructose-bisphosphatase, Zinc-Free, from Rabbit Liver. In: Methods in enzymology. 1982 ; Vol. 90, No. C. pp. 327-329.
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Demaine, MM, Caperelli, CA & Benkovic, S 1982, 'Fructose-bisphosphatase, Zinc-Free, from Rabbit Liver', Methods in enzymology, vol. 90, no. C, pp. 327-329. https://doi.org/10.1016/S0076-6879(82)90149-5

Fructose-bisphosphatase, Zinc-Free, from Rabbit Liver. / Demaine, Margaret M.; Caperelli, Carol A.; Benkovic, Stephen.

In: Methods in enzymology, Vol. 90, No. C, 01.01.1982, p. 327-329.

Research output: Contribution to journalArticle

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