Distances between Structural Metal Ion, Substrates, and Allosteric Modifier of Fructose Bisphosphatase

Brian A. Cunningham, Frank M. Raushel, Joseph J. Villafranca, Stephen J. Benkovic

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The binding of two paramagnetic probes within a subunit of fructose bisphosphatase, viz., Mn2+ at a structural site and a nitroxide spin-label at a sulfhydryl site, has permitted the measurement of NMR and electron paramagnetic resonance (EPR) relaxation rates to map the active and allosteric site topography. Distances from these loci to the phosphoryl of fructose 6-phosphate (Fru-6-P) and inorganic phosphate (Pi) and four nuclei of adenosine 5'-phosphate (AMP) (the phosphorus nucleus, H-8, FI-2, and H-V) were obtained. These measurements located the Mn2+ approximately equidistant from the two phosphoryl moieties of the product ligands Fru-6-P and Pi and in close proximity to the AMP. The adenosine moiety of the latter is oriented anti. Analysis of EPR data revealed that the nitroxide group is ~16Å from the structural Mn2+ site. Calculation of the residence times for the hydrolysis reaction products suggests that their dissociation should not be rate limiting in the overall reaction cycle.

Original languageEnglish (US)
Pages (from-to)359-362
Number of pages4
JournalBiochemistry
Volume20
Issue number2
DOIs
StatePublished - Jan 1981

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Structural metals
Fructose-Bisphosphatase
Adenosine Monophosphate
Metal ions
Metals
Electron Spin Resonance Spectroscopy
Ions
Paramagnetic resonance
Substrates
Allosteric Site
Spin Labels
Reaction products
Adenosine
Phosphorus
Topography
Hydrolysis
Catalytic Domain
Phosphates
Nuclear magnetic resonance
Ligands

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Cunningham, B. A., Raushel, F. M., Villafranca, J. J., & Benkovic, S. J. (1981). Distances between Structural Metal Ion, Substrates, and Allosteric Modifier of Fructose Bisphosphatase. Biochemistry, 20(2), 359-362. https://doi.org/10.1021/bi00505a020
Cunningham, Brian A. ; Raushel, Frank M. ; Villafranca, Joseph J. ; Benkovic, Stephen J. / Distances between Structural Metal Ion, Substrates, and Allosteric Modifier of Fructose Bisphosphatase. In: Biochemistry. 1981 ; Vol. 20, No. 2. pp. 359-362.
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Cunningham, BA, Raushel, FM, Villafranca, JJ & Benkovic, SJ 1981, 'Distances between Structural Metal Ion, Substrates, and Allosteric Modifier of Fructose Bisphosphatase', Biochemistry, vol. 20, no. 2, pp. 359-362. https://doi.org/10.1021/bi00505a020

Distances between Structural Metal Ion, Substrates, and Allosteric Modifier of Fructose Bisphosphatase. / Cunningham, Brian A.; Raushel, Frank M.; Villafranca, Joseph J.; Benkovic, Stephen J.

In: Biochemistry, Vol. 20, No. 2, 01.1981, p. 359-362.

Research output: Contribution to journalArticle

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