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Structural properties of an active form of rabbit muscle phosphofructokinase.

Publication ,  Journal Article
Hesterberg, LK; Lee, JC; Erickson, HP
Published in: J Biol Chem
September 25, 1981

The quaternary structure of an active form of rabbit muscle phosphofructokinase was studied by sedimentation and electron microscopy. Active enzyme centrifugation studies at pH 7.0 and 23 +/- 1 degrees C showed that phosphofructokinase sediments as a single component with a sedimentation coefficient of 12.2 +/- 0.5 S. Identical results were obtained in two assay and three solvent systems. Boundary sedimentation studies of phosphofructokinase in the presence of 1.0 mM fructose 6-phosphate, 0.1 mM adenylyl imidodiphosphate at pH 7.0 and 23 +/- 1 degrees C were performed. The results showed that the sedimentation coefficient of phosphofructokinase remains constant within the range of protein concentration studied and assumes a value of 12.4 S. The molecular weights of the subunit and the 12.4 S component were measured by sedimentation equilibrium yielding values of 83,000 and 330,000 for the monomeric and polymeric species, respectively. It is, therefore, concluded that the active form of phosphofructokinase is indeed the tetrameric species. The structure of the phosphofructokinase tetramer was also studied by electron microscopy of negatively stained specimens. Particles identified as tetramers measured approximately 9 nm in diameter by 14 nm in length. The observed size and shape are consistent with the hydrodynamic measurements. Structural features within the tetramer were interpreted as due to the four individual subunits, each one approximately 4 X 6 X 6 nm in size, arranged with D2 symmetry.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

September 25, 1981

Volume

256

Issue

18

Start / End Page

9724 / 9730

Location

United States

Related Subject Headings

  • Rabbits
  • Protein Conformation
  • Phosphofructokinase-1
  • Muscles
  • Models, Molecular
  • Microscopy, Electron
  • Macromolecular Substances
  • Kinetics
  • Biochemistry & Molecular Biology
  • Animals
 

Citation

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Hesterberg, L. K., Lee, J. C., & Erickson, H. P. (1981). Structural properties of an active form of rabbit muscle phosphofructokinase. J Biol Chem, 256(18), 9724–9730.
Hesterberg, L. K., J. C. Lee, and H. P. Erickson. “Structural properties of an active form of rabbit muscle phosphofructokinase.J Biol Chem 256, no. 18 (September 25, 1981): 9724–30.
Hesterberg LK, Lee JC, Erickson HP. Structural properties of an active form of rabbit muscle phosphofructokinase. J Biol Chem. 1981 Sep 25;256(18):9724–30.
Hesterberg, L. K., et al. “Structural properties of an active form of rabbit muscle phosphofructokinase.J Biol Chem, vol. 256, no. 18, Sept. 1981, pp. 9724–30.
Hesterberg LK, Lee JC, Erickson HP. Structural properties of an active form of rabbit muscle phosphofructokinase. J Biol Chem. 1981 Sep 25;256(18):9724–9730.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

September 25, 1981

Volume

256

Issue

18

Start / End Page

9724 / 9730

Location

United States

Related Subject Headings

  • Rabbits
  • Protein Conformation
  • Phosphofructokinase-1
  • Muscles
  • Models, Molecular
  • Microscopy, Electron
  • Macromolecular Substances
  • Kinetics
  • Biochemistry & Molecular Biology
  • Animals