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. 1974 Jun;139(3):665–675. doi: 10.1042/bj1390665

The preparation and properties of pyruvate kinase from yeast

David A Fell 1,*, Peter F Liddle 1,†, Arthur R Peacocke 1,‡, Raymond A Dwek 2
PMCID: PMC1166331  PMID: 4369339

Abstract

A new method is described for the preparation of pyruvate kinase from yeast. This eliminates proteolysis during the preparation. The molecular weight of yeast pyruvate kinase is 215000, and it is composed of four subunits. Such properties of the enzyme as its extinction coefficient, cold-lability, thiol-group reactivity and binding of Mn2+ ions are compared with those previously reported for yeast pyruvate kinase prepared by different methods. The specific activity is significantly higher than previously observed, but otherwise the enzyme is similar, apart from its molecular weight and Mn2+-binding characteristics, to preparations from Saccharomyces cerevisiae obtained in this laboratory (e.g. Fell et al., 1972, and references therein) and that of C. H. Suelter (e.g. Kuczenski & Suelter, 1971, and references therein), and is different from the enzyme isolated from Saccharomyces carlsbergensis by B. Hess and his co-workers (e.g. Wieker & Hess, 1972, and references therein).

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Selected References

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

  1. Ainsworth S., MacFarlane N. A kinetic study of rabbit muscle pyruvate kinase. Biochem J. 1973 Feb;131(2):223–236. doi: 10.1042/bj1310223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashton Karen, Peacocke A. R. The molecular weight of a yeast pyruvate kinase and its subunits. FEBS Lett. 1971 Jul 15;16(1):25–28. doi: 10.1016/0014-5793(71)80675-0. [DOI] [PubMed] [Google Scholar]
  3. Bischofberger H., Hess B., Röschlau P. Molecular weight of yeast pyruvate kinase. Hoppe Seylers Z Physiol Chem. 1971 Aug;352(8):1139–1150. doi: 10.1515/bchm2.1971.352.2.1139. [DOI] [PubMed] [Google Scholar]
  4. Bischofberger H., Hess B., Röschlau P., Wieker H. J., Zimmermann-Telschow H. Amino-acid composition and subunit structure of yeast-pyruvate kinase. Hoppe Seylers Z Physiol Chem. 1970 Mar;351(3):401–408. doi: 10.1515/bchm2.1970.351.1.401. [DOI] [PubMed] [Google Scholar]
  5. Bornmann L., Röschlau P., Hess B. Essentielle Aminosäurereste von Hefe-Pyruvatkinase. Hoppe Seylers Z Physiol Chem. 1972 May;353(5):696–696. [PubMed] [Google Scholar]
  6. Chervenka C. H. Long-column meniscus depletion sedimentation equilibrium technique for the analytical ultracentrifuge. Anal Biochem. 1970 Mar;34:24–29. doi: 10.1016/0003-2697(70)90082-5. [DOI] [PubMed] [Google Scholar]
  7. Cottam G. L., Mildvan A. S., Hunsley J. R., Suelter C. H. Proton relaxation and kinetic studies of ternary complexes of an allosteric pyruvate kinase from yeast. J Biol Chem. 1972 Jun 25;247(12):3802–3809. [PubMed] [Google Scholar]
  8. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  9. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  10. Fell D. A., Peacocke A. R., Dwek R. A. Proton-relaxation-enhancement studies on the binding to yeast pyruvate kinase of a substrate and effectors. Eur J Biochem. 1972 Aug 18;29(1):128–133. doi: 10.1111/j.1432-1033.1972.tb01966.x. [DOI] [PubMed] [Google Scholar]
  11. Flashner M., Hollenberg P. F., Coon M. J. Mechanism of action of pyruvate kinase. Role of sulfhydryl groups in catalytic activity as determined by disulfide interchange. J Biol Chem. 1972 Dec 25;247(24):8114–8121. [PubMed] [Google Scholar]
  12. Foord R., Jakeman E., Oliver C. J., Pike E. R., Blagrove R. J., Wood E., Peacocke A. R. Determination of diffusion coefficients of haemocyanin at low concentration by intensity fluctuation spectroscopy of scattered laser light. Nature. 1970 Jul 18;227(5255):242–245. doi: 10.1038/227242a0. [DOI] [PubMed] [Google Scholar]
  13. Haeckel R., Hess B., Lauterborn W., Wüster K. H. Purification and allosteric properties of yeast pyruvate kinase. Hoppe Seylers Z Physiol Chem. 1968 May;349(5):699–714. doi: 10.1515/bchm2.1968.349.1.699. [DOI] [PubMed] [Google Scholar]
  14. Hunsley J. R., Suelter C. H. Yeast pyruvate kinase. I. Purification and some chemical properties. J Biol Chem. 1969 Sep 25;244(18):4815–4818. [PubMed] [Google Scholar]
  15. Hunsley J. R., Suelter C. H. Yeast pyruvate kinase. II. Kinetic properties. J Biol Chem. 1969 Sep 25;244(18):4819–4822. [PubMed] [Google Scholar]
  16. ITZHAKI R. F., GILL D. M. A MICRO-BIURET METHOD FOR ESTIMATING PROTEINS. Anal Biochem. 1964 Dec;9:401–410. doi: 10.1016/0003-2697(64)90200-3. [DOI] [PubMed] [Google Scholar]
  17. Kayne F. J., Price N. C. Conformational changes in the allosteric inhibition of muscle pyruvate kinase by phenylalanine. Biochemistry. 1972 Nov 7;11(23):4415–4420. doi: 10.1021/bi00773a031. [DOI] [PubMed] [Google Scholar]
  18. Kuczenski R. T., Suelter C. H. Effect of temperature and effectors on the conformations of yeast pyruvate kinase. Biochemistry. 1970 Feb 17;9(4):939–945. doi: 10.1021/bi00806a033. [DOI] [PubMed] [Google Scholar]
  19. Kuczenski R. T., Suelter C. H. Fructose 1,6-diphosphate enhances inactivation of yeast pyruvate kinase at 23. Evidence for a stabilized dimer intermediate. Biochemistry. 1971 Jul 20;10(15):2867–2872. doi: 10.1021/bi00791a011. [DOI] [PubMed] [Google Scholar]
  20. Kuczenski R. T., Suelter C. H. Yeast pyruvate kinase. Native and subunit molecular weight. Biochemistry. 1970 May 12;9(10):2043–2047. doi: 10.1021/bi00812a003. [DOI] [PubMed] [Google Scholar]
  21. LENNEY J. F. A study of two yeast proteinases. J Biol Chem. 1956 Aug;221(2):919–930. [PubMed] [Google Scholar]
  22. Levitzki A., Stallcup W. B., Koshland D. E., Jr Half-of-the-sites reactivity and the conformational states of cytidine triphosphate synthetase. Biochemistry. 1971 Aug 31;10(18):3371–3378. doi: 10.1021/bi00794a009. [DOI] [PubMed] [Google Scholar]
  23. Macfarlane N., Ainsworth S. A kinetic study of Baker's-yeast pyruvate kinase activated by fructose 1,6-diphosphate. Biochem J. 1972 Oct;129(5):1035–1047. doi: 10.1042/bj1291035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Röschlau P., Hess B. Purification and crystallization of yeast pyruvate kinase. Hoppe Seylers Z Physiol Chem. 1972 Mar;353(3):435–440. doi: 10.1515/bchm2.1972.353.1.435. [DOI] [PubMed] [Google Scholar]
  25. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  26. VANHOLDE K. E., COHEN L. B. PHYSICAL STUDIES OF HEMOCYANINS. I. CHARACTERIZATION AND SUBUNIT STRUCTURE OF LOLIGO PEALEI HEMOCYANIN. Biochemistry. 1964 Dec;3:1803–1808. doi: 10.1021/bi00900a001. [DOI] [PubMed] [Google Scholar]
  27. Wieker H. J., Hess B. Function of thiol groups in yeast pyruvate kinase. Hoppe Seylers Z Physiol Chem. 1972 Dec;353(12):1877–1893. doi: 10.1515/bchm2.1972.353.2.1877. [DOI] [PubMed] [Google Scholar]

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