Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Mar;63(3):956–961. doi: 10.1128/aem.63.3.956-961.1997

Identification of ATP-dependent phosphofructokinase as a regulatory step in the glycolytic pathway of the actinomycete Streptomyces coelicolor A3(2).

A M Alves 1, G J Euverink 1, M J Bibb 1, L Dijkhuizen 1
PMCID: PMC168387  PMID: 9055413

Abstract

The ATP-dependent phosphofructokinase (ATP-PFK) of Streptomyces coelicolor A3(2) was purified to homogeneity (1,600-fold) and characterized (110 kDa, with a single type of subunit of 40 kDa); it is allosterically inhibited by phosphoenolpyruvate. Cloning of the pfk gene of S. coelicolor A3(2) and analysis of the deduced amino acid sequence (343 amino acids; 36,667 Da) revealed high similarities to the PPi-PFK enzyme from Amycolatopsis methanolica (tetramer, nonallosteric; 70%) and to the allosteric ATP-PFK enzymes from other bacteria, e.g., Escherichia coli (tetramer; 37%) and Bacillus stearothermophilus (tetramer, 41%). Further structural and functional analysis of the two actinomycete PFK enzymes should elucidate the features of these proteins that determine substrate specificity (ATP versus PPi) and allosteric (in)sensitivity.

Full Text

The Full Text of this article is available as a PDF (182.7 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Alves A. M., Euverink G. J., Hektor H. J., Hessels G. I., van der Vlag J., Vrijbloed J. W., Hondmann D., Visser J., Dijkhuizen L. Enzymes of glucose and methanol metabolism in the actinomycete Amycolatopsis methanolica. J Bacteriol. 1994 Nov;176(22):6827–6835. doi: 10.1128/jb.176.22.6827-6835.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alves A. M., Meijer W. G., Vrijbloed J. W., Dijkhuizen L. Characterization and phylogeny of the pfp gene of Amycolatopsis methanolica encoding PPi-dependent phosphofructokinase. J Bacteriol. 1996 Jan;178(1):149–155. doi: 10.1128/jb.178.1.149-155.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Angell S., Lewis C. G., Buttner M. J., Bibb M. J. Glucose repression in Streptomyces coelicolor A3(2): a likely regulatory role for glucose kinase. Mol Gen Genet. 1994 Jul 25;244(2):135–143. doi: 10.1007/BF00283514. [DOI] [PubMed] [Google Scholar]
  5. Auzat I., Le Bras G., Branny P., De La Torre F., Theunissen B., Garel J. R. The role of Glu187 in the regulation of phosphofructokinase by phosphoenolpyruvate. J Mol Biol. 1994 Jan 7;235(1):68–72. doi: 10.1016/s0022-2836(05)80014-2. [DOI] [PubMed] [Google Scholar]
  6. Branny P., De La Torre F., Garel J. R. Cloning, sequencing, and expression in Escherichia coli of the gene coding for phosphofructokinase in Lactobacillus bulgaricus. J Bacteriol. 1993 Sep;175(17):5344–5349. doi: 10.1128/jb.175.17.5344-5349.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carlisle S. M., Blakeley S. D., Hemmingsen S. M., Trevanion S. J., Hiyoshi T., Kruger N. J., Dennis D. T. Pyrophosphate-dependent phosphofructokinase. Conservation of protein sequence between the alpha- and beta-subunits and with the ATP-dependent phosphofructokinase. J Biol Chem. 1990 Oct 25;265(30):18366–18371. [PubMed] [Google Scholar]
  8. Chevalier C., Saillard C., Bové J. M. Organization and nucleotide sequences of the Spiroplasma citri genes for ribosomal protein S2, elongation factor Ts, spiralin, phosphofructokinase, pyruvate kinase, and an unidentified protein. J Bacteriol. 1990 May;172(5):2693–2703. doi: 10.1128/jb.172.5.2693-2703.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Currie P. D., Sullivan D. T. Structure and expression of the gene encoding phosphofructokinase (PFK) in Drosophila melanogaster. J Biol Chem. 1994 Oct 7;269(40):24679–24687. [PubMed] [Google Scholar]
  10. Fothergill-Gilmore L. A., Michels P. A. Evolution of glycolysis. Prog Biophys Mol Biol. 1993;59(2):105–235. doi: 10.1016/0079-6107(93)90001-z. [DOI] [PubMed] [Google Scholar]
  11. Green P. C., Latshaw S. P., Ladror U. S., Kemp R. G. Identification of critical lysyl residues in the pyrophosphate-dependent phosphofructo-1-kinase of Propionibacterium freudenreichii. Biochemistry. 1992 May 26;31(20):4815–4821. doi: 10.1021/bi00135a011. [DOI] [PubMed] [Google Scholar]
  12. Heinisch J. Isolation and characterization of the two structural genes coding for phosphofructokinase in yeast. Mol Gen Genet. 1986 Jan;202(1):75–82. doi: 10.1007/BF00330520. [DOI] [PubMed] [Google Scholar]
  13. Hellinga H. W., Evans P. R. Nucleotide sequence and high-level expression of the major Escherichia coli phosphofructokinase. Eur J Biochem. 1985 Jun 3;149(2):363–373. doi: 10.1111/j.1432-1033.1985.tb08934.x. [DOI] [PubMed] [Google Scholar]
  14. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  15. Huang M., Albach R. A., Chang K. P., Tripathi R. L., Kemp R. G. Cloning and sequencing a putative pyrophosphate-dependent phosphofructokinase gene from Entamoeba histolytica. Biochim Biophys Acta. 1995 Jan 25;1260(2):215–217. doi: 10.1016/0167-4781(94)00216-p. [DOI] [PubMed] [Google Scholar]
  16. Kemp R. G., Tripathi R. L. Pyrophosphate-dependent phosphofructo-1-kinase complements fructose 1,6-bisphosphatase but not phosphofructokinase deficiency in Escherichia coli. J Bacteriol. 1993 Sep;175(17):5723–5724. doi: 10.1128/jb.175.17.5723-5724.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kieser T., Hopwood D. A. Genetic manipulation of Streptomyces: integrating vectors and gene replacement. Methods Enzymol. 1991;204:430–458. doi: 10.1016/0076-6879(91)04023-h. [DOI] [PubMed] [Google Scholar]
  18. Klein R. D., Olson E. R., Favreau M. A., Winterrowd C. A., Hatzenbuhler N. T., Shea M. H., Nulf S. C., Geary T. G. Cloning of a cDNA encoding phosphofructokinase from Haemonchus contortus. Mol Biochem Parasitol. 1991 Sep;48(1):17–26. doi: 10.1016/0166-6851(91)90160-8. [DOI] [PubMed] [Google Scholar]
  19. Kotlarz D., Buc H. Two Escherichia coli fructose-6-phosphate kinases. Preparative purification, oligomeric structure and immunological studies. Biochim Biophys Acta. 1977 Sep 15;484(1):35–48. doi: 10.1016/0005-2744(77)90111-5. [DOI] [PubMed] [Google Scholar]
  20. Ladror U. S., Gollapudi L., Tripathi R. L., Latshaw S. P., Kemp R. G. Cloning, sequencing, and expression of pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii. J Biol Chem. 1991 Sep 5;266(25):16550–16555. [PubMed] [Google Scholar]
  21. Lee C. P., Kao M. C., French B. A., Putney S. D., Chang S. H. The rabbit muscle phosphofructokinase gene. Implications for protein structure, function, and tissue specificity. J Biol Chem. 1987 Mar 25;262(9):4195–4199. [PubMed] [Google Scholar]
  22. Llanos R. M., Harris C. J., Hillier A. J., Davidson B. E. Identification of a novel operon in Lactococcus lactis encoding three enzymes for lactic acid synthesis: phosphofructokinase, pyruvate kinase, and lactate dehydrogenase. J Bacteriol. 1993 May;175(9):2541–2551. doi: 10.1128/jb.175.9.2541-2551.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nakajima H., Noguchi T., Yamasaki T., Kono N., Tanaka T., Tarui S. Cloning of human muscle phosphofructokinase cDNA. FEBS Lett. 1987 Oct 19;223(1):113–116. doi: 10.1016/0014-5793(87)80519-7. [DOI] [PubMed] [Google Scholar]
  24. O'Brien W. E., Bowien S., Wood H. G. Isolation and characterization of a pyrophosphate-dependent phosphofructokinase from Propionibacterium shermanii. J Biol Chem. 1975 Nov 25;250(22):8690–8695. [PubMed] [Google Scholar]
  25. Plunkett G., 3rd, Burland V., Daniels D. L., Blattner F. R. Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes. Nucleic Acids Res. 1993 Jul 25;21(15):3391–3398. doi: 10.1093/nar/21.15.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Poolman B., Smid E. J., Veldkamp H., Konings W. N. Bioenergetic consequences of lactose starvation for continuously cultured Streptococcus cremoris. J Bacteriol. 1987 Apr;169(4):1460–1468. doi: 10.1128/jb.169.4.1460-1468.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Redenbach M., Kieser H. M., Denapaite D., Eichner A., Cullum J., Kinashi H., Hopwood D. A. A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol. 1996 Jul;21(1):77–96. doi: 10.1046/j.1365-2958.1996.6191336.x. [DOI] [PubMed] [Google Scholar]
  28. Reeves R. E., South D. J., Blytt H. J., Warren L. G. Pyrophosphate:D-fructose 6-phosphate 1-phosphotransferase. A new enzyme with the glycolytic function of 6-phosphofructokinase. J Biol Chem. 1974 Dec 25;249(24):7737–7741. [PubMed] [Google Scholar]
  29. Rozario C., Smith M. W., Müller M. Primary sequence of a putative pyrophosphate-linked phosphofructokinase gene of Giardia lamblia. Biochim Biophys Acta. 1995 Jan 25;1260(2):218–222. doi: 10.1016/0167-4781(94)00217-q. [DOI] [PubMed] [Google Scholar]
  30. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  31. Sakai H., Ohta T. Molecular cloning and nucleotide sequence of the gene for pyruvate kinase of Bacillus stearothermophilus and the production of the enzyme in Escherichia coli. Evidence that the genes for phosphofructokinase and pyruvate kinase constitute an operon. Eur J Biochem. 1993 Feb 1;211(3):851–859. doi: 10.1111/j.1432-1033.1993.tb17618.x. [DOI] [PubMed] [Google Scholar]
  32. Shirakihara Y., Evans P. R. Crystal structure of the complex of phosphofructokinase from Escherichia coli with its reaction products. J Mol Biol. 1988 Dec 20;204(4):973–994. doi: 10.1016/0022-2836(88)90056-3. [DOI] [PubMed] [Google Scholar]
  33. Takahashi N., Kalfas S., Yamada T. Phosphorylating enzymes involved in glucose fermentation of Actinomyces naeslundii. J Bacteriol. 1995 Oct;177(20):5806–5811. doi: 10.1128/jb.177.20.5806-5811.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Uyeda K. Phosphofructokinase. Adv Enzymol Relat Areas Mol Biol. 1979;48:193–244. doi: 10.1002/9780470122938.ch4. [DOI] [PubMed] [Google Scholar]
  36. Valdez B. C., Chang S. H., Younathan E. S. Site-directed mutagenesis at the regulatory site of fructose 6-phosphate-1-kinase from Bacillus stearothermophilus. Biochem Biophys Res Commun. 1988 Oct 14;156(1):537–542. doi: 10.1016/s0006-291x(88)80875-1. [DOI] [PubMed] [Google Scholar]
  37. Wessberg K. L., Skolnick S., Xu J., Marciano-Cabral F., Kemp R. G. Cloning, sequencing and expression of the pyrophosphate-dependent phosphofructo-1-kinase from Naegleria fowleri. Biochem J. 1995 Apr 1;307(Pt 1):143–149. doi: 10.1042/bj3070143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wright F., Bibb M. J. Codon usage in the G+C-rich Streptomyces genome. Gene. 1992 Apr 1;113(1):55–65. doi: 10.1016/0378-1119(92)90669-g. [DOI] [PubMed] [Google Scholar]
  39. Xu J., Seki M., Denda K., Yoshida M. Molecular cloning of phosphofructokinase 1 gene from a thermophilic bacterium, Thermus thermophilus. Biochem Biophys Res Commun. 1991 May 15;176(3):1313–1318. doi: 10.1016/0006-291x(91)90429-b. [DOI] [PubMed] [Google Scholar]
  40. van Alebeek G. J., Keltjens J. T., van der Drift C. Purification and characterization of inorganic pyrophosphatase from Methanobacterium thermoautotrophicum (strain delta H). Biochim Biophys Acta. 1994 Jun 12;1206(2):231–239. doi: 10.1016/0167-4838(94)90213-5. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES