, 11:647 | Cite as

Characterization of cofactor-dependent and cofactor-independent phosphoglycerate mutases from Archaea

Original Paper


Phosphoglycerate mutases (PGM) catalyze the reversible conversion of 3-phosphoglycerate and 2-phosphoglycerate as part of glycolysis and gluconeogenesis. Two structural and mechanistically unrelated types of PGMs are known, a cofactor (2,3-bisphosphoglycerate)-dependent (dPGM) and a cofactor-independent enzyme (iPGM). Here, we report the characterization of the first archaeal cofactor-dependent PGM from Thermoplasma acidophilum, which is encoded by ORF TA1347. This ORF was cloned and expressed in Escherichia coli and the recombinant protein was characterized as functional dPGM. The enzyme constitutes a 46 kDa homodimeric protein. Enzyme activity required 2,3-bisphosphoglycerate as cofactor and was inhibited by vanadate, a specific inhibitor of dPGMs in bacteria and eukarya; inhibition could be partially relieved by EDTA. Histidine 23 of the archaeal dPGM of T. acidophilum, which corresponds to active site histidine in dPGMs from bacteria and eukarya, was exchanged for alanine by site directed mutagenesis. The H23A mutant was catalytically inactive supporting the essential role of H23 in catalysis of the archaeal dPGM. Further, an archaeal cofactor-independent PGM encoded by ORF AF1751 from the hyperthermophilic sulfate reducer Archaeoglobus fulgidus was characterized after expression in E. coli. The monomeric 46 kDa protein showed cofactor-independent PGM activity and was stimulated by Mn2+ and exhibited high thermostability up to 70°C. A comprehensive phylogenetic analysis of both types of archaeal phosphoglycerate mutases is also presented.


Phosphoglycerate mutases Archaea Thermoplasma acidophilum Archaeoglobus fulgidus 


  1. Bond CS, White MF, Hunter WN (2001) High resolution structure of the phosphohistidine-activated form of Escherichia coli cofactor-dependent phosphoglycerate mutase. J Biol Chem 276:3247–3253PubMedCrossRefGoogle Scholar
  2. Bond CS, White MF, Hunter WN (2002) Mechanistic implications for Escherichia coli cofactor-dependent phosphoglycerate mutase based on the high-resolution crystal structure of a vanadate complex. J Mol Biol 316:1071–1081PubMedCrossRefGoogle Scholar
  3. Carreras J, Bartrons R, Grisolia S (1980) Vanadate inhibits 2,3-bisphosphoglycerate dependent phosphoglycerate mutases but does not affect the 2,3-bisphosphoglycerate independent phosphoglycerate mutases. Biochem Biophys Res Commun 96:1267–1273PubMedGoogle Scholar
  4. Collet JF, Stroobant V, Van Schaftingen E (2001) The 2,3-bisphosphoglycerate-independent phosphoglycerate mutase from Trypanosoma brucei: metal-ion dependency and phosphoenzyme formation. FEMS Microbiol Lett 204:39–44PubMedCrossRefGoogle Scholar
  5. Deppenmeier U, Johann A, Hartsch T, Merkl R, Schmitz RA, Martinez-Arias R, Henne A, Wiezer A, Baumer S, Jacobi C, Bruggemann H, Lienard T, Christmann A, Bomeke M, Steckel S, Bhattacharyya A, Lykidis A, Overbeek R, Klenk HP, Gunsalus RP, Fritz HJ, Gottschalk G (2002) The genome of Methanosarcina mazei: evidence for lateral gene transfer between bacteria and archaea. J Mol Microbiol Biotechnol 4:453–461PubMedGoogle Scholar
  6. Fothergill-Gilmore LA, Watson HC (1989) The phosphoglycerate mutases. Adv Enzymol Relat Areas Mol Biol 62:227–313PubMedCrossRefGoogle Scholar
  7. Fraser HI, Kvaratskhelia M, White MF (1999) The two analogous phosphoglycerate mutases of Escherichia coli. FEBS Lett 455:344–348PubMedCrossRefGoogle Scholar
  8. Galperin MY, Jedrzejas MJ (2001) Conserved core structure and active site residues in alkaline phosphatase superfamily enzymes. Proteins 45:318–324PubMedCrossRefGoogle Scholar
  9. Galperin MY, Bairoch A, Koonin EV (1998) A superfamily of metalloenzymes unifies phosphopentomutase and cofactor-independent phosphoglycerate mutase with alkaline phosphatases and sulfatases. Prot Sci 7:1829–1835Google Scholar
  10. Gouet P, Courcelle E, Stuart DI, Metoz F (1999) ESPript: analysis of multiple sequence alignments in PostScript. Bioinformatics 15:305–308PubMedCrossRefGoogle Scholar
  11. Graham DE, Xu H, White RH (2002) A divergent archaeal member of the alkaline phosphatase binuclear metalloenzyme superfamily has phosphoglycerate mutase activity. FEBS Lett 517:190–194PubMedCrossRefGoogle Scholar
  12. Grana X, Perez dlO, Broceno C, Stocker M, Garriga J, Puigdomenech P, Climent F (1995) 2,3-Bisphosphoglycerate-independent phosphoglycerate mutase is conserved among different phylogenic kingdoms. Comp Biochem Physiol B Biochem Mol Biol 112:287–293PubMedCrossRefGoogle Scholar
  13. Guerra DG, Vertommen D, Fothergill-Gilmore LA, Opperdoes FR, Michels PA (2004) Characterization of the cofactor-independent phosphoglycerate mutase from Leishmania mexicana mexicana. Histidines that coordinate the two metal ions in the active site show different susceptibilities to irreversible chemical modification. Eur J Biochem 271:1798–1810PubMedCrossRefGoogle Scholar
  14. Jedrzejas MJ (2000) Structure, function, and evolution of phosphoglycerate mutases: comparison with fructose-2,6-bisphosphatase, acid phosphatase, and alkaline phosphatase. Prog Biophys Mol Biol 73:263–287PubMedCrossRefGoogle Scholar
  15. Jedrzejas MJ, Chander M, Setlow P, Krishnasamy G (2000) Structure and mechanism of action of a novel phosphoglycerate mutase from Bacillus stearothermophilus. EMBO J 19:1419–1431PubMedCrossRefGoogle Scholar
  16. Jones DT (1999) Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol 292:195–202PubMedCrossRefGoogle Scholar
  17. Koonin EV, Mushegian AR, Galperin MY, Walker DR (1997) Comparison of archaeal and bacterial genomes: computer analysis of protein sequences predicts novel functions and suggests a chimeric origin for the archaea. Mol Microbiol 25:619–637PubMedCrossRefGoogle Scholar
  18. Möller-Zinkhan D, Thauer RK (1990) Anaerobic lactate oxidation to 3 CO2 by Archaeoglobus fulgidus via the carbon monoxide dehydrogenase pathway: demonstration of the acetyl-CoA carbon–carbon cleavage reaction in cell extracts. Arch Microbiol 153:215–218CrossRefGoogle Scholar
  19. Nelson KE, Clayton RA, Gill SR, Gwinn ML, Dodson RJ, Haft DH, Hickey EK, Peterson JD, Nelson WC, Ketchum KA, McDonald L, Utterback TR, Malek JA, Linher KD, Garrett MM, Stewart AM, Cotton MD, Pratt MS, Phillips CA, Richardson D, Heidelberg J, Sutton GG, Fleischmann RD, Eisen JA, Fraser CM (1999) Evidence for lateral gene transfer between Archaea and bacteria from genome sequence of Thermotoga maritima. Nature 399:323–329PubMedCrossRefGoogle Scholar
  20. Pearson CL, Loshon CA, Pedersen LB, Setlow B, Setlow P (2000) Analysis of the function of a putative 2,3-diphosphoglyceric acid-dependent phosphoglycerate mutase from Bacillus subtilis. J Bacteriol 182:4121–4123PubMedCrossRefGoogle Scholar
  21. Potters MB, Solow BT, Bischoff KM, Graham DE, Lower BH, Helm R, Kennelly PJ (2003) Phosphoprotein with Phosphoglycerate mutase activity from the Archaeon Sulfolobus solfataricus. J Bacteriol 185:2112–2121PubMedCrossRefGoogle Scholar
  22. Price NC, Jaenicke R (1982) The quaternary structure of phosphoglycerate mutase from yeast: evidence against dissociation of the tetrameric enzyme at low concentrations. FEBS Lett 143:283–286PubMedCrossRefGoogle Scholar
  23. Rigden DJ, Bagyan I, Lamani E, Setlow P, Jedrzejas MJ (2001) A cofactor-dependent phosphoglycerate mutase homolog from Bacillus stearothermophilus is actually a broad specificity phosphatase. Prot Sci 10:1835–1846CrossRefGoogle Scholar
  24. Ronimus RS, Morgan HW (2003) Distribution and phylogenies of enzymes of the Embden–Meyerhof–Parnas pathway from archaea and hyperthermophilic bacteria support a gluconeogenic origin of metabolism. Archaea 1:199–221PubMedCrossRefGoogle Scholar
  25. Rose ZB (1971) The phosphorylation of yeast phosphoglycerate mutase. Arch Biochem Biophys 146:359–360PubMedCrossRefGoogle Scholar
  26. Saavedra E, Encalada R, Pineda E, Jasso-Chavez R, Moreno-Sanchez R (2005) Glycolysis in Entamoeba histolytica. Biochemical characterization of recombinant glycolytic enzymes and flux control analysis. FEBS J 272:1767–1783PubMedCrossRefGoogle Scholar
  27. Selkov E, Maltsev N, Olsen GJ, Overbeek R, Whitman WB (1997) A reconstruction of the metabolism of Methanococcus jannaschii from sequence data. Gene 197:GC11–GC26PubMedCrossRefGoogle Scholar
  28. Shima S, Sordel-Klippert M, Brioukhanov A, Netrusov A, Linder D, Thauer RK (2001) Characterization of a heme-dependent catalase from Methanobrevibacter arboriphilus. Appl Environ Microbiol 67:3041–3045PubMedCrossRefGoogle Scholar
  29. Siebers B, Schönheit P (2005) Unusual pathways and enzymes of central carbohydrate metabolism in Archaea. Curr Opin Microbiol 8:695–705PubMedGoogle Scholar
  30. Stetter KO (1988) Archaeoglobus fulgidus gen. nov., sp. nov.: a new taxon of extremely thermophilic archaebacteria. Syst Appl Microbiol 10:172–173Google Scholar
  31. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  32. Towne JC, Rodwell VW, Grisolia S (1957) The microestimation, distribution, and biosynthesis of 2,3-diphosphoglyceric acid. J Biol Chem 226:777–788PubMedGoogle Scholar
  33. Uhrinova S, Uhrin D, Nairn J, Price NC, Fothergill-Gilmore LA, Barlow PN (2001) Solution structure and dynamics of an open beta-sheet, glycolytic enzyme, monomeric 23.7 kDa phosphoglycerate mutase from Schizosaccharomyces pombe. J Mol Biol 306:275–290PubMedCrossRefGoogle Scholar
  34. Van der Oost J, Huynen MA, Verhees CH (2002) Molecular characterization of phosphoglycerate mutase in archaea. FEMS Microbiol Lett 212:111–120PubMedCrossRefGoogle Scholar
  35. White MF, Fothergill-Gilmore LA (1992) Development of a mutagenesis, expression and purification system for yeast phosphoglycerate mutase. Investigation of the role of active-site His181. Eur J Biochem 207:709–714PubMedCrossRefGoogle Scholar
  36. Zhang Y, Foster JM, Kumar S, Fougere M, Carlow CK (2004) Cofactor-independent phosphoglycerate mutase has an essential role in Caenorhabditis elegans and is conserved in parasitic nematodes. J Biol Chem 279:37185–37190PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2007

Authors and Affiliations

  1. 1.Institut für Allgemeine MikrobiologieChristian-Albrechts-Universität KielKielGermany

Personalised recommendations