1887

Abstract

SUMMARY: Polyphosphate kinase, an enzyme which incorporated the -phosphate of ATP into long-chain polyphosphate molecules, was purified more than 700-fold from by ammonium sulphate fractionation, DEAE-cellulose column chromatography and Sephadex G-200 gel filtration. The enzyme had a broad pH optimum at 6·0 to 7·0 and required Mn or Mg, histone, and inorganic phosphate for activity. The for Mn-ATP was 0·53 m, and for inorganic phosphate was 1·67 m.

Free ATP concentrations greater than 8 inhibited the enzyme. Free Mn or Mg concentrations greater than 2 m or 6 m, respectively, were also inhibitory. Activity was strongly inhibited by 4 m-ADP, 1 m-PP or 20 m-NaF. The effect of ADP might have resulted from reversing the equilibrium of the kinase reaction. The activation by phosphate ions might indicate a role for the enzyme in regulating intracellular phosphate levels or maintaining a phosphorus reserve. The level of enzymic activity in the bacteria responded to changes in inorganic phosphate concentration in the medium. Basic proteins, such as protamine, could substitute for histone as activator. Proteins such as casein or bovine serum albumin would also substitute for histone but only in the absence of inorganic phosphate. The presence of a protein might be necessary to form a complex with the product, thus preventing reversal of the reaction

The reaction product was characterized, and found to be labile in hydroxylamine, base, and acid at 100 °C. It behaved as a long-chain-polyphosphate molecule on chromatography in an Ebel’s solvent. The enzymic activity was therefore not that of a protein kinase.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-88-1-65
1975-05-01
2021-05-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/88/1/mic-88-1-65.html?itemId=/content/journal/micro/10.1099/00221287-88-1-65&mimeType=html&fmt=ahah

References

  1. Agabian N., Rosen O. M., Shapiro L. 1972; Characterization of a protein acyl kinase from Caulobacter crescentus. Biochemical and Biophysical Research Communications 49:1690–1698
    [Google Scholar]
  2. Baker A. L., Schmidt R. R. 1964; Induced utilization of polyphosphate during nuclear division in synchronously growing Chlorella. Biochimica et biophysica acta 93:180–182
    [Google Scholar]
  3. Bonner J., Chalkley G. R., Dahmus M., Fambrough D., Fujimura F., Huang R. C., Huberman J., Jensen R., Marushige K., Ohlenbusch H., Olivera B., Widholm J. 1968; Isolation and characterization of chromosomal nucleoproteins. Methods in Enzymology 12B New York: Academic Press;
    [Google Scholar]
  4. Harold F. M. 1966; Inorganic polyphosphates in biology: structure, metabolism, and function. Bacteriological Reviews 30:772–794
    [Google Scholar]
  5. Hokin L. E., Sastry P. S., Galsworthy P. R., Yoda A. 1965; Evidence that a phosphorylated intermediate in a brain transport adenosine triphosphatase is an acyl phosphate. Proceedings of the National Academy of Sciences of the United States of America 54:177–184
    [Google Scholar]
  6. Kabat D. 1970; Phosphorylation of ribosomal proteins in rabbit reticulocytes. Characterization and regulatory aspects. Biochemistry 9:4160–4175
    [Google Scholar]
  7. Kaltwasser H. 1962; Die Rolle der Polyphosphate im Phosphatstoffwechsel eines Knallgasbakteriums (Hydrogenomonas Stamm o). Archiv für Mikrobiologie 41:282–306
    [Google Scholar]
  8. Kornberg S. R. 1957; Adenosine triphosphate synthesis from polyphosphate by an enzyme from Escherichia coli. Biochimica et biophysica acta 26:294–300
    [Google Scholar]
  9. Kornberg A., Kornberg S. R., Simms E. S. 1956; Metaphosphate synthesis by an enzyme from Escherichia coli. Biochimica et biophysica acta 26:215–227
    [Google Scholar]
  10. Krulwich T. A., Ensign J. C., Tipper D. J., Strominger J. L. 1967; Sphere-rod morphogenesis in Arthrobacter crystallopoietes. II. Peptides of the cell wall peptidoglycan. Journal of Bacteriology 94:741–750
    [Google Scholar]
  11. Kundig W., Roseman S. 1971; Sugar transport. I. Isolation of a phosphotransferase system from Escherichia coli. Journal of Biological Chemistry 246:1393–1406
    [Google Scholar]
  12. Li H.-C., Brown G. G. 1973; Orthophosphate and histone dependent polyphosphate kinase from E. coli. Biochemical and Biophysical Research Communications 53:875–881
    [Google Scholar]
  13. Li H.-C., Felmly D. A. 1973; A rapid paper chromatographic assay for protein kinase. Analytical Biochemistry 52:300–304
    [Google Scholar]
  14. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  15. Martonosi A. 1969; Sarcoplasmic reticulum. VII. Properties of a phosphoprotein intermediate implicated in calcium transport. Journal of Biological Chemistry 244:613–620
    [Google Scholar]
  16. Mcconnell D. J., Bonner J. 1972; Preparation of highly purified ribonucleic acid polymerase; separation from polynucleotide phosphorylase and polyphosphate kinase. Biochemistry 11:4329–4336
    [Google Scholar]
  17. Miyachi S., Kanai R., Mihara S., Miyachi S., Aoki S. 1964; Metabolic roles of inorganic polyphosphates in Chlorella cells. Biochimica et biophysica acta 93:625–634
    [Google Scholar]
  18. Mudd S., Yoshida A., Koike M. 1958; Polyphosphate as accumulator of phosphorus and energy. Journal of Bacteriology 75:224–235
    [Google Scholar]
  19. Mühlradt P. F. 1971; Synthesis of high molecular weight polyphosphate with a partially purified enzyme from Salmonella. Journal of General Microbiology 68:115–122
    [Google Scholar]
  20. Nishi A. 1961; Role of polyphosphate and phospholipid in germinating spores of Aspergillus niger. Journal of Bacteriology 81:10–19
    [Google Scholar]
  21. Ohashi S., Van Wazer J. R. 1963; Paper chromatography of very long chain polyphosphates. Analytical Chemistry 35:1984–1985
    [Google Scholar]
  22. Schechter S. L., Li H.-C., Krulwich T. A. 1972; Regulation of inducible enzyme synthesis in Arthrobacter atrocyaneus. Abstracts of the Annual Meetings of the American Society for Microbiology p. 176:
    [Google Scholar]
  23. Walaas E. 1958; Stability constants of metal complexes with mononucleotides. Acta chemica scandinavica 12:528–536
    [Google Scholar]
  24. Winder F. G., Denneny J. M. 1957; The metabolism of inorganic polyphosphate in mycobacteria. Journal of General Microbiology 17:573–585
    [Google Scholar]
  25. Yoshida A., Yamataka A. 1953; On the metaphosphate of the yeast. Journal of Biochemistry 40:85–94
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-88-1-65
Loading
/content/journal/micro/10.1099/00221287-88-1-65
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error