Studies on structure of human erythrocyte phosphofructokinase

N. S. Karadsheh, K. Uyeda, R. M. Oliver

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Abstract

Human erythrocyte phosphofructokinase was purified to homogeneity and some of its physicochemical properties were investigated. The enzyme exists in several polymeric states. At high protein concentrations (3 mg/ml), its predominant form shows a sedimentation coefficient of 57 S. In highly dilute solution (0.3 to 10 μg/ml), the enzyme is in 12 S to 18 S forms as shown by an 'active enzyme' centrifugation technique. Phosphofructokinase dissociates in 6 M guanidinium/HCl to a subunit the molecular weight of which is 80,000 as determined by high speed sedimentation equilibrium. Polyacrylamide gel electrophoresis in S.D.S. however, reveals 2 protein bands of which the m.w. was 80,000 and 85,000. The 85,000-dalton subunit is similar to the single subunit which occurs in human muscle phosphofructokinase. The 80,000-dalton subunit is designated as the human erythrocyte enzyme subunit. The subunits were separated by DEAE-cellulose chromatography in the presence of 8 M urea, and appeared to occur in the ratio of 1:2.9 (muscle:erythrocyte) based upon a comparison of the relative intensity of the stained protein bands on acrylamide gels. The amino acid composition and the tryptic peptide map of human erythrocyte phosphofructokinase are different from those of human muscle phosphofructokinase. Electron microscopic observations show 2 dominant sets of images. One has a polyhedronal shape which is probably an aggregate of erythrocyte type of subunits. The other aggregate form, which is similar to those of human and rabbit muscle phosphofructokinases, is suggested to be the polymers of muscle type subunits of human erythrocyte phosphofructokinase.

Original languageEnglish (US)
Pages (from-to)3515-3524
Number of pages10
JournalJournal of Biological Chemistry
Volume252
Issue number10
StatePublished - Jan 1 1977

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Karadsheh, N. S., Uyeda, K., & Oliver, R. M. (1977). Studies on structure of human erythrocyte phosphofructokinase. Journal of Biological Chemistry, 252(10), 3515-3524.