Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1974 Mar;137(3):463–467. doi: 10.1042/bj1370463

Preparation and properties of a phosphoenzyme from the phosphoglucomutase of Micrococcus lysodeikticus

John B Clarke 1, Michael Birch 1, Hubert G Britton 1
PMCID: PMC1166145  PMID: 4472509

Abstract

1. Phosphoglucomutase from Micrococcus lysodeikticus was incubated with 14C- and 32P-labelled glucose 1,6-diphosphate and separated from the cofactor on a Sephadex column. 32P-labelled phosphate (0.7mol/mol of enzyme) was associated with the enzyme, but no 14C label was. 2. The 32P-labelled enzyme exchanged its label with the substrates. When the labelled enzyme was incubated in Tris buffer, pH8.3, at 30°C the proportion of exchangeable label slowly fell indicating a half-life of the phosphoenzyme of about 50h. 3. When HClO4 was added to the labelled phosphoenzyme all of the label was precipitated with the protein and none was released as Pi. On alkaline hydrolysis Pi was released at a rate comparable with the rate of hydrolysis of the phosphoenzyme from rabbit muscle. 4. We conclude that the phosphoenzyme from Micrococcus lysodeikticus yields a relatively stable, catalytically active phosphoenzyme when treated with cofactor, and that there is no evidence for the formation of an enzyme–glucose 1,6-diphosphate complex. The properties of the phosphoenzyme, which resemble those of rabbit muscle phosphoglucomutase, suggest that the phosphate may be bound to serine.

Full text

PDF
463

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ANDERSON L., JOLLES G. R. A study of the linkage of phosphorus to protein in phosphoglucomutase. Arch Biochem Biophys. 1957 Jul;70(1):121–128. doi: 10.1016/0003-9861(57)90085-1. [DOI] [PubMed] [Google Scholar]
  2. Britton H. G., Clarke J. B. The mechanism of the phosphoglucomutase reaction. Studies on rabbit muscle phosphoglucomutase with flux techniques. Biochem J. 1968 Nov;110(2):161–180. doi: 10.1042/bj1100161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clarke J. B., Britton H. G. The mechanism of phosphoglucomutase from Micrococcus lysodeikticus. Biochem J. 1974 Mar;137(3):453–461. doi: 10.1042/bj1370453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunstan P. M., Anthony C., Drabble W. T. Microbial metabolism of C 1 and C 2 compounds. The involvement of glycollate in the metabolism of ethanol and of acetate by Pseudomonas AM1. Biochem J. 1972 Jun;128(1):99–106. doi: 10.1042/bj1280099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hanabusa K., Dougherty H. W., Del Río C., Hashimoto T., Handler P. Phosphoglucomutase. II. Preparation and properties of phosphoglucomutases from Micrococcuus lysodeikticus and Bacillus cereus. J Biol Chem. 1966 Sep 10;241(17):3930–3939. [PubMed] [Google Scholar]
  6. Hashimoto T., Handler P. Phosphoglucomutase. 3. Purification and properties of phosphoglucomutases from flounder and shark muscle. J Biol Chem. 1966 Sep 10;241(17):3940–3948. [PubMed] [Google Scholar]
  7. JOSHI J. G., HANDLER P. PHOSPHOGLUCOMUTASE. I. PURIFICATION AND PROPERTIES OF PHOSPHOGLUCOMUTASE FROM ESCHERICHIA COLI. J Biol Chem. 1964 Sep;239:2741–2751. [PubMed] [Google Scholar]
  8. KENNEDY E. P., KOSHLAND D. E., Jr Properties of the phosphorylated active site of phosphoglucomutase. J Biol Chem. 1957 Sep;228(1):419–431. [PubMed] [Google Scholar]
  9. Marshall R. D., Zamecnik P. C. Some physical properties of lysyl and arginyl-transfer RNA synthetases of Escherichia coli B. Biochim Biophys Acta. 1969 Jul 1;181(2):454–464. doi: 10.1016/0005-2795(69)90279-7. [DOI] [PubMed] [Google Scholar]
  10. Milstein C. P., Clarke J. B., Britton H. G. The reactive serine residue in phosphoglucomutase of Micrococcus lysodeikiticus. Biochem J. 1973 Nov;135(3):551–553. doi: 10.1042/bj1350551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. RAY W. J., Jr, ROSCELLI G. A. A KINETIC STUDY OF THE PHOSPHOGLUCOMUTASE PATHWAY. J Biol Chem. 1964 Apr;239:1228–1236. [PubMed] [Google Scholar]
  12. RAY W. J., Jr, ROSCELLI G. A. THE PHOSPHOGLUCOMUTASE PATHWAY. AN INVESTIGATION OF PHOSPHO-ENZYME ISOMERIZATION. J Biol Chem. 1964 Nov;239:3935–3941. [PubMed] [Google Scholar]
  13. SUGINO Y., MIYOSHI Y. THE SPECIFIC PRECIPITATION OF ORTHOPHOSPHATE AND SOME BIOCHEMICAL APPLICATIONS. J Biol Chem. 1964 Jul;239:2360–2364. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES