Studies of the structure of fructose-6-phosphate 2-kinase: Fructose-2,6-bisphosphatase

Ryuzo Sakakibara, Tatsuya Tanaka, Kosaku Uyeda, E. Glen Richards, Helmut Thomas, Kenji Kangawa, Hisayuki Matsuo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Some physicochemical properties of a homogeneous preparation of a bifunctional enzyme, fructose-6-phosphate 2-kinase:fructose-2,6-bisphosphatase, were determined. The molecular weight of the enzyme is 101 000 as determined by high-speed sedimentation equilibrium. The molecular weight of dissociated enzyme is 55 000 in 6 M guanidinium chloride by sedimentation equilibrium and in sodium dodecyl sulfate by polyacrylamide gel electrophoresis. A value of 4.7 was observed for the isoelectric point. Tryptic peptide maps and high-performance liquid chromatography of the trypsin-digested enzyme revealed approximately 60 peptides. Amino acid analysis of the enzyme shows that it contains 27 lysine and 36 arginine residues per 55 000 daltons. No free N-terminal amino acid residue was detectable, suggesting that it is blocked. Hydrolysis of the enzyme by carboxypeptidases A and B releases tyrosine followed by histidine and arginine, indicating that the amino acid sequence at the carboxyl terminus is probably -Arg-His-Tyr. Tryptic digestion of [32P]phosphofructose-6-phosphate 2-kinase:fructose-2,6-bisphosphatase yields a 32P-labeled peptide detected by tryptic peptide mapping and high-performance liquid chromatography. Thermolysin digestion of CNBr-cleaved 32P-enzyme also yields a single 32P-peptide. These results indicate that fructose-6-phosphate 2-kinase:fructose-2,6-bisphosphatase is a dimer of 55 000 daltons and the subunits are very similar, if not identical.

Original languageEnglish (US)
Pages (from-to)6818-6824
Number of pages7
JournalBiochemistry
Volume24
Issue number24
StatePublished - 1985

Fingerprint

Phosphofructokinase-2
Enzymes
Peptides
High performance liquid chromatography
Sedimentation
Amino Acids
Arginine
Digestion
Molecular Weight
Molecular weight
High Pressure Liquid Chromatography
Carboxypeptidase B
Carboxypeptidases A
Thermolysin
Peptide Mapping
Guanidine
Isoelectric Point
Electrophoresis
Histidine
Sodium Dodecyl Sulfate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sakakibara, R., Tanaka, T., Uyeda, K., Richards, E. G., Thomas, H., Kangawa, K., & Matsuo, H. (1985). Studies of the structure of fructose-6-phosphate 2-kinase: Fructose-2,6-bisphosphatase. Biochemistry, 24(24), 6818-6824.

Studies of the structure of fructose-6-phosphate 2-kinase : Fructose-2,6-bisphosphatase. / Sakakibara, Ryuzo; Tanaka, Tatsuya; Uyeda, Kosaku; Richards, E. Glen; Thomas, Helmut; Kangawa, Kenji; Matsuo, Hisayuki.

In: Biochemistry, Vol. 24, No. 24, 1985, p. 6818-6824.

Research output: Contribution to journalArticle

Sakakibara, R, Tanaka, T, Uyeda, K, Richards, EG, Thomas, H, Kangawa, K & Matsuo, H 1985, 'Studies of the structure of fructose-6-phosphate 2-kinase: Fructose-2,6-bisphosphatase', Biochemistry, vol. 24, no. 24, pp. 6818-6824.
Sakakibara R, Tanaka T, Uyeda K, Richards EG, Thomas H, Kangawa K et al. Studies of the structure of fructose-6-phosphate 2-kinase: Fructose-2,6-bisphosphatase. Biochemistry. 1985;24(24):6818-6824.
Sakakibara, Ryuzo ; Tanaka, Tatsuya ; Uyeda, Kosaku ; Richards, E. Glen ; Thomas, Helmut ; Kangawa, Kenji ; Matsuo, Hisayuki. / Studies of the structure of fructose-6-phosphate 2-kinase : Fructose-2,6-bisphosphatase. In: Biochemistry. 1985 ; Vol. 24, No. 24. pp. 6818-6824.
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