Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1999 Jan 1;337(Pt 1):89–95.

The role of the C-terminal region in phosphoglycerate mutase.

R A Walter 1, J Nairn 1, D Duncan 1, N C Price 1, S M Kelly 1, D J Rigden 1, L A Fothergill-Gilmore 1
PMCID: PMC1219940  PMID: 9854029

Abstract

Removal of the C-terminal seven residues from phosphoglycerate mutase from Saccharomyces cerevisiae by limited proteolysis is associated with loss of mutase activity, but no change in phosphatase activity. The presence of the cofactor 2, 3-bisphosphoglycerate, or of the cofactor and substrate 3-phosphoglycerate together, confers protection against proteolysis. The substrate alone offers no protection. Replacement of either or both of the two lysines at the C-terminus by glycines has only limited effects on the kinetic properties of phosphoglycerate mutase, indicating that these residues are unlikely to be involved in crucial electrostatic interactions with the substrate, intermediate or product in the reaction. However, the double-mutant form of the enzyme is more sensitive to proteolysis and is no longer protected against proteolysis by the presence of cofactor. The proteolysed wild-type and two of the mutated forms of the enzyme show a reduced response to 2-phosphoglycollate, which enhances the instability of the phospho form of the native enzyme. The phosphoglycerate mutase from Schizosaccharomyces pombe, which lacks the analogous C-terminal tail, has an inherently lower mutase activity and is also less responsive to stimulation by 2-phosphoglycollate. It is proposed that the C-terminal region of phosphoglycerate mutase helps to maintain the enzyme in its active phosphorylated form and assists in the retention of the bisphosphoglycerate intermediate at the active site. However, its role seems not to be to contribute directly to ligand binding, but rather to exert indirect effects on the transfer of the phospho group between substrate, enzyme, intermediate and product.

Full Text

The Full Text of this article is available as a PDF (175.2 KB).

Selected References

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

  1. Blouquit Y., Calvin M. C., Rosa R., Promé D., Promé J. C., Pratbernou F., Cohen-Solal M., Rosa J. Sequence of the human erythrocyte phosphoglycerate mutase by microsequencer and mass spectrometry. J Biol Chem. 1988 Nov 15;263(32):16906–16910. [PubMed] [Google Scholar]
  2. Castellà-Escolà J., Montoliu L., Pons G., Puigdomènech P., Cohen-Solal M., Carreras J., Rigau J., Climent F. Sequence of rat skeletal muscle phosphoglycerate mutase cDNA. Biochem Biophys Res Commun. 1989 Dec 29;165(3):1345–1351. doi: 10.1016/0006-291x(89)92751-4. [DOI] [PubMed] [Google Scholar]
  3. Currie P. D., Sullivan D. T. Structure, expression and duplication of genes which encode phosphoglyceromutase of Drosophila melanogaster. Genetics. 1994 Oct;138(2):352–363. [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. EDELHOCH H., RODWELL V. W., GRISOLIA S. The molecular properties of yeast and muscle phosphoglyceric acid mutase. J Biol Chem. 1957 Oct;228(2):891–903. [PubMed] [Google Scholar]
  6. Fleischmann R. D., Adams M. D., White O., Clayton R. A., Kirkness E. F., Kerlavage A. R., Bult C. J., Tomb J. F., Dougherty B. A., Merrick J. M. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995 Jul 28;269(5223):496–512. doi: 10.1126/science.7542800. [DOI] [PubMed] [Google Scholar]
  7. Fothergill-Gilmore L. A., Watson H. C. The phosphoglycerate mutases. Adv Enzymol Relat Areas Mol Biol. 1989;62:227–313. doi: 10.1002/9780470123089.ch6. [DOI] [PubMed] [Google Scholar]
  8. Garel M. C., Joulin V., Le Boulch P., Calvin M. C., Préhu M. O., Arous N., Longin R., Rosa R., Rosa J., Cohen-Solal M. Human bisphosphoglycerate mutase. Expression in Escherichia coli and use of site-directed mutagenesis in the evaluation of the role of the carboxyl-terminal region in the enzymatic mechanism. J Biol Chem. 1989 Nov 15;264(32):18966–18972. [PubMed] [Google Scholar]
  9. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  10. Girg R., Rudolph R., Jaenicke R. Limited proteolysis of porcine-muscle lactic dehydrogenase by thermolysin during reconstitution yields dimers. Eur J Biochem. 1981 Oct;119(2):301–305. doi: 10.1111/j.1432-1033.1981.tb05608.x. [DOI] [PubMed] [Google Scholar]
  11. Hayes J. D., Kerr L. A., Cronshaw A. D. Evidence that glutathione S-transferases B1B1 and B2B2 are the products of separate genes and that their expression in human liver is subject to inter-individual variation. Molecular relationships between the B1 and B2 subunits and other Alpha class glutathione S-transferases. Biochem J. 1989 Dec 1;264(2):437–445. doi: 10.1042/bj2640437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson C. M., Price N. C. Denaturation and renaturation of the monomeric phosphoglycerate mutase from Schizosaccharomyces pombe. Biochem J. 1987 Jul 15;245(2):525–530. doi: 10.1042/bj2450525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joulin V., Peduzzi J., Roméo P. H., Rosa R., Valentin C., Dubart A., Lapeyre B., Blouquit Y., Garel M. C., Goossens M. Molecular cloning and sequencing of the human erythrocyte 2,3-bisphosphoglycerate mutase cDNA: revised amino acid sequence. EMBO J. 1986 Sep;5(9):2275–2283. doi: 10.1002/j.1460-2075.1986.tb04495.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Le Boulch P., Joulin V., Garel M. C., Rosa J., Cohen-Solal M. Molecular cloning and nucleotide sequence of murine 2,3-bisphosphoglycerate mutase cDNA. Biochem Biophys Res Commun. 1988 Oct 31;156(2):874–881. doi: 10.1016/s0006-291x(88)80925-2. [DOI] [PubMed] [Google Scholar]
  16. Li L., Ling S., Wu C. l., Yao W., Xu G. Separate bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase: the role of the C-terminal tail in modulating enzyme activity. Biochem J. 1997 Dec 15;328(Pt 3):751–756. doi: 10.1042/bj3280751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Liu S., Gresser M. J., Tracey A. S. 1H and 51V NMR studies of the interaction of vanadate and 2-vanadio-3-phosphoglycerate with phosphoglycerate mutase. Biochemistry. 1992 Mar 17;31(10):2677–2685. doi: 10.1021/bi00125a007. [DOI] [PubMed] [Google Scholar]
  18. Nairn J., Krell T., Coggins J. R., Pitt A. R., Fothergill-Gilmore L. A., Walter R., Price N. C. The use of mass spectrometry to examine the formation and hydrolysis of the phosphorylated form of phosphoglycerate mutase. FEBS Lett. 1995 Feb 13;359(2-3):192–194. doi: 10.1016/0014-5793(95)00044-a. [DOI] [PubMed] [Google Scholar]
  19. Nairn J., Price N. C., Fothergill-Gilmore L. A., Walker G. E., Fothergill J. E., Dunbar B. The amino acid sequence of the small monomeric phosphoglycerate mutase from the fission yeast Schizosaccharomyces pombe. Biochem J. 1994 Feb 1;297(Pt 3):603–608. doi: 10.1042/bj2970603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nairn J., Price N. C., Kelly S. M., Rigden D., Fothergill-Gilmore L. A., Krell T. Phosphoglycerate mutase from Schizosaccharomyces pombe: development of an expression system and characterisation of three histidine mutants of the enzyme. Biochim Biophys Acta. 1996 Aug 15;1296(1):69–75. doi: 10.1016/0167-4838(96)00046-5. [DOI] [PubMed] [Google Scholar]
  21. Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., Inada T., Itoh T., Kajihara M. A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12.7-28.0 min region on the linkage map. DNA Res. 1996 Jun 30;3(3):137–155. doi: 10.1093/dnares/3.3.137. [DOI] [PubMed] [Google Scholar]
  22. Price N. C., Duncan D., McAlister J. W. Inactivation of rabbit muscle phosphoglycerate mutase by limited proteolysis with thermolysin. Biochem J. 1985 Jul 1;229(1):167–171. doi: 10.1042/bj2290167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rigden D. J., Alexeev D., Phillips S. E., Fothergill-Gilmore L. A. The 2.3 A X-ray crystal structure of S. cerevisiae phosphoglycerate mutase. J Mol Biol. 1998 Feb 20;276(2):449–459. doi: 10.1006/jmbi.1997.1554. [DOI] [PubMed] [Google Scholar]
  24. Rose Z. B., Dube S. Rates of phosphorylation and dephosphorylation of phosphoglycerate mutase and bisphosphoglycerate synthase. J Biol Chem. 1976 Aug 25;251(16):4817–4822. [PubMed] [Google Scholar]
  25. Rost B., Sander C. Combining evolutionary information and neural networks to predict protein secondary structure. Proteins. 1994 May;19(1):55–72. doi: 10.1002/prot.340190108. [DOI] [PubMed] [Google Scholar]
  26. Rost B., Sander C. Prediction of protein secondary structure at better than 70% accuracy. J Mol Biol. 1993 Jul 20;232(2):584–599. doi: 10.1006/jmbi.1993.1413. [DOI] [PubMed] [Google Scholar]
  27. Rost B., Sander C., Schneider R. PHD--an automatic mail server for protein secondary structure prediction. Comput Appl Biosci. 1994 Feb;10(1):53–60. doi: 10.1093/bioinformatics/10.1.53. [DOI] [PubMed] [Google Scholar]
  28. Sakoda S., Shanske S., DiMauro S., Schon E. A. Isolation of a cDNA encoding the B isozyme of human phosphoglycerate mutase (PGAM) and characterization of the PGAM gene family. J Biol Chem. 1988 Nov 15;263(32):16899–16905. [PubMed] [Google Scholar]
  29. Sasaki R., Hirose M., Sugimoto E., Chiba H. Studies on a role of the 2,3-diphosphoglycerate phosphatase activity in the yeast phosphoglycerate mutase reaction. Biochim Biophys Acta. 1971 Mar 10;227(3):595–607. doi: 10.1016/0005-2744(71)90010-6. [DOI] [PubMed] [Google Scholar]
  30. Sasaki R., Sugimoto R., Chiba H. Yeast phosphoglyceric acid mutase-modifying enzyme. Arch Biochem Biophys. 1966 Jul;115(1):53–61. doi: 10.1016/s0003-9861(66)81037-8. [DOI] [PubMed] [Google Scholar]
  31. Sedmak J. J., Grossberg S. E. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. doi: 10.1016/0003-2697(77)90428-6. [DOI] [PubMed] [Google Scholar]
  32. Shanske S., Sakoda S., Hermodson M. A., DiMauro S., Schon E. A. Isolation of a cDNA encoding the muscle-specific subunit of human phosphoglycerate mutase. J Biol Chem. 1987 Oct 25;262(30):14612–14617. [PubMed] [Google Scholar]
  33. Ureña J. M., Graña X., de Lecea L., Ruiz P., Castellà J., Carreras J., Pons G., Climent F. Isolation and sequencing of a cDNA encoding the B isozyme of rat phosphoglycerate mutase. Gene. 1992 Apr 15;113(2):281–282. doi: 10.1016/0378-1119(92)90408-h. [DOI] [PubMed] [Google Scholar]
  34. White M. F., Fothergill-Gilmore L. A. Development of a mutagenesis, expression and purification system for yeast phosphoglycerate mutase. Investigation of the role of active-site His181. Eur J Biochem. 1992 Jul 15;207(2):709–714. doi: 10.1111/j.1432-1033.1992.tb17099.x. [DOI] [PubMed] [Google Scholar]
  35. White M. F., Fothergill-Gilmore L. A. Sequence of the gene encoding phosphoglycerate mutase from Saccharomyces cerevisiae. FEBS Lett. 1988 Mar 14;229(2):383–387. doi: 10.1016/0014-5793(88)81161-x. [DOI] [PubMed] [Google Scholar]
  36. White P. J., Nairn J., Price N. C., Nimmo H. G., Coggins J. R., Hunter I. S. Phosphoglycerate mutase from Streptomyces coelicolor A3(2): purification and characterization of the enzyme and cloning and sequence analysis of the gene. J Bacteriol. 1992 Jan;174(2):434–440. doi: 10.1128/jb.174.2.434-440.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Winn S. I., Watson H. C., Harkins R. N., Fothergill L. A. Structure and activity of phosphoglycerate mutase. Philos Trans R Soc Lond B Biol Sci. 1981 Jun 26;293(1063):121–130. doi: 10.1098/rstb.1981.0066. [DOI] [PubMed] [Google Scholar]
  38. Yanagawa S., Hitomi K., Sasaki R., Chiba H. Isolation and characterization of cDNA encoding rabbit reticulocyte 2,3-bisphosphoglycerate synthase. Gene. 1986;44(2-3):185–191. doi: 10.1016/0378-1119(86)90181-2. [DOI] [PubMed] [Google Scholar]
  39. Yomano L. P., Scopes R. K., Ingram L. O. Cloning, sequencing, and expression of the Zymomonas mobilis phosphoglycerate mutase gene (pgm) in Escherichia coli. J Bacteriol. 1993 Jul;175(13):3926–3933. doi: 10.1128/jb.175.13.3926-3933.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES