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. 1972 Oct;112(1):327–336. doi: 10.1128/jb.112.1.327-336.1972

Purification and Properties of the Adenosine Diphosphate-Glucose and Uridine Diphosphate-Glucose Pyrophosphorylases of Mycobacterium smegmatis: Inhibition and Activation of the Adenosine Diphosphate-Glucose Pyrophosphorylase

David Lapp a,1, Alan D Elbein a
PMCID: PMC251415  PMID: 5079067

Abstract

Crude extracts of Mycobacterium smegmatis catalyzed the synthesis of adenosine diphosphate-glucose (ADP-Glc), cytidine diphosphate-glucose, guanosine diphosphate-glucose (GDP-Glc), thymidine diphosphate-glucose (TDP-Glc), and uridine diphosphate-glucose (UDP-Glc). In these crude enzyme fractions, high concentrations of trehalose-P inhibited the ADP-Glc and GDP-Glc pyrophosphorylases but did not effect the UDP-Glc or TDP-Glc pyrophosphorylases. Both the ADP-Glc pyrophosphorylase and the UDP-Glc pyrophosphorylase were partially purified (about 140-fold and 60-fold, respectively), and their properties were compared. For the ADP-Glc pyrophosphorylase, the Km for adenosine triphosphate was 6 × 10−4m, whereas that for glucose-1-P was 8 × 10−4m. The optimal concentration of Mg2+ was 1 × 10−3m, and the pH optimum was 8.5. For the UDP-Glc pyrophosphorylase, the Km for uridine triphosphate was 1 × 10−3m and for glucose-1-P was 2 × 10−3m. The optimal Mg2+ concentration was 1 × 10−3m, and the pH optimum was about 8.0. The purified ADP-Glc pyrophosphorylase was inhibited by fructose-6-P, fructose-1, 6-diphosphate, glucose-6-P, and phosphoenolpyruvate. On the other hand, trehalose, trehalose diphosphate, sodium pyruvate, and ribose-5-P did not effect the ADP-Glc pyrophosphorylase. None of these compounds, including trehalose-P, had any effect on the UDP-Glc pyrophosphorylase.

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Selected References

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

  1. Albrecht G. J., Bass S. T., Seifert L. L., Hansen R. G. Crystallization and properties of uridine diphosphate glucose pyrophosphorylase from liver. J Biol Chem. 1966 Jun 25;241(12):2968–2975. [PubMed] [Google Scholar]
  2. Antoine A. D., Tepper B. S. Characterization of glycogens from mycobacteria. Arch Biochem Biophys. 1969 Oct;134(1):207–213. doi: 10.1016/0003-9861(69)90267-7. [DOI] [PubMed] [Google Scholar]
  3. Antoine A. D., Tepper B. S. Environmental control of glycogen and lipid content of Mycobacterium phlei. J Gen Microbiol. 1969 Feb;55(2):217–226. doi: 10.1099/00221287-55-2-217. [DOI] [PubMed] [Google Scholar]
  4. BERNSTEIN R. L., ROBBINS P. W. CONTROL ASPECTS OF URIDINE 5'-DIPHOSPHATE GLUCOSE AND THYMIDINE 5'-DIPHOSPHATE GLUCOSE SYNTHESIS BY MICROBIAL ENZYMES. J Biol Chem. 1965 Jan;240:391–397. [PubMed] [Google Scholar]
  5. Ghosh H. P., Preiss J. Adenosine diphosphate glucose pyrophosphorylase. A regulatory enzyme in the biosynthesis of starch in spinach leaf chloroplasts. J Biol Chem. 1966 Oct 10;241(19):4491–4504. [PubMed] [Google Scholar]
  6. Kornfeld R. H., Ginsburg V. Control of synthesis of guanosine 5'-diphosphate D-mannose and guanosine 5'-diphosphate L-fucose in bacteria. Biochim Biophys Acta. 1966 Mar 28;117(1):79–87. doi: 10.1016/0304-4165(66)90154-1. [DOI] [PubMed] [Google Scholar]
  7. Liu C., Patterson B. W., Lapp D., Elbein A. D. Trehalose phosphate synthesis from uridine diphosphate glucose or guanosine diphosphate glucose. Activation of uridine diphosphate-glucose: trehalose phosphate synthetase by polynucleotides. J Biol Chem. 1969 Jul 10;244(13):3728–3731. [PubMed] [Google Scholar]
  8. MACDONALD D. L., WONG R. Y. A CHEMICAL SYNTHESIS OF TREHALOSE 6-PHOSPHATE. Biochim Biophys Acta. 1964 May 11;86:390–392. doi: 10.1016/0304-4165(64)90066-2. [DOI] [PubMed] [Google Scholar]
  9. MAYER R. M., GINSBURG V. PURIFICATION AND PROPERTIES OF CYTIDINE DIPHOSPHATE D-GLUCOSE PYROPHOSPHORYLASE FROM SALMONELLA PARATYPHI A. J Biol Chem. 1965 May;240:1900–1904. [PubMed] [Google Scholar]
  10. MELO A., GLASER L. THE NUCLEOTIDE SPECIFICITY AND FEEDBACK CONTROL OF THYMIDINE DIPHOSPHATE D-GLUCOSE PYROPHOSPHORYLASE. J Biol Chem. 1965 Jan;240:398–405. [PubMed] [Google Scholar]
  11. Matula M., Mitchell M., Elbein A. D. Partial purification and properties of a highly specific trehalose phosphate phosphatase from Mycobacterium smegmatis. J Bacteriol. 1971 Jul;107(1):217–222. doi: 10.1128/jb.107.1.217-222.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nakae T., Nikaido H. Multiple molecular forms of uridine diphosphate glucose pyrophosphorylase from Salmonella typhimurium. I. Catalytic properties of various forms. J Biol Chem. 1971 Jul 25;246(14):4386–4396. [PubMed] [Google Scholar]
  13. SHEN L., PREISS J. BIOSYNTHESIS OF BACTERIAL GLYCOGEN. I. PURIFICATION AND PROPERTIES OF THE ADENOSINE DIPHOSPHOGLUCOSE PYROPHOSPHORYLASE OF ARTHROBACTER SPECIES NRRL B1973. J Biol Chem. 1965 Jun;240:2334–2340. [PubMed] [Google Scholar]
  14. Sanwal G. G., Preiss J. Biosynthesis of starch in Chlorella pyrenoidosa. II. Regulation of ATP: alpha-D-glucose 1-phosphate adenyl transferase (ADP-glucose pyrophosphorylase) by inorganic phosphate and 3-phosphoglycerate. Arch Biochem Biophys. 1967 Mar;119(1):454–469. doi: 10.1016/0003-9861(67)90477-8. [DOI] [PubMed] [Google Scholar]
  15. Shen L. C., Atkinson D. E. Regulation of adenosine diphosphate glucose synthase from Escherichia coli. Interactions of adenylate energy charge and modifier concentrations. J Biol Chem. 1970 Aug 10;245(15):3996–4000. [PubMed] [Google Scholar]
  16. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  17. Tsuboi K. K., Fukunaga K., Petricciani J. C. Purification and specific kinetic properties of erythrocyte uridine diphosphate glucose pyrophosphorylase. J Biol Chem. 1969 Feb 10;244(3):1008–1015. [PubMed] [Google Scholar]
  18. VILLAR-PALASI C., LARNER J. Uridinediphosphate glucose pyrophosphorylase from skeletal muscle. Arch Biochem Biophys. 1960 Jan;86:61–66. doi: 10.1016/0003-9861(60)90368-4. [DOI] [PubMed] [Google Scholar]
  19. WINDER F., BRENNAN P. THE ACCUMULATION OF FREE TREHALOSE BY MYCOBACTERIA EXPOSED TO ISONIAZID. Biochim Biophys Acta. 1964 Aug 19;90:442–444. doi: 10.1016/0304-4165(64)90222-3. [DOI] [PubMed] [Google Scholar]
  20. Winder F. G., Rooney S. A. The effects of isoniazid on the carbohydrates of Mycobacterium tuberculosis BCG. Biochem J. 1970 Apr;117(2):355–368. doi: 10.1042/bj1170355. [DOI] [PMC free article] [PubMed] [Google Scholar]

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