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. 1970 Jun;118(2):253–258. doi: 10.1042/bj1180253

Purification and properties of the nicotinamide–adenine dinucleotide phosphate-dependent isocitrate dehydrogenase from pig liver cytoplasm

J A Illingworth 1, K F Tipton 1
PMCID: PMC1179111  PMID: 4394948

Abstract

The NADP-dependent isocitrate dehydrogenase from pig liver soluble fraction was purified over 500-fold with an overall yield of 25%. The purified enzyme, which is homogeneous by all the usual criteria, has a molecular weight of about 75000 and is composed of two identical subunits. This has been demonstrated by ultracentrifugation, fluorescence titration and peptide `fingerprinting'. The maximal turnover number, extinction coefficients at 280nm and 260nm and amino acid analysis are described.

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Selected References

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

  1. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CHEN R. F., PLAUT G. W. ACTIVATION AND INHIBITION OF DPN-LINKED ISOCITRATE DEHYDROGENASE OF HEART BY CERTAIN NUCLEOTIDES. Biochemistry. 1963 Sep-Oct;2:1023–1032. doi: 10.1021/bi00905a020. [DOI] [PubMed] [Google Scholar]
  3. Colman R. F. Binding of substrates by native and chemically modified isocitrate dehydrogenase. Biochim Biophys Acta. 1969 Nov 4;191(2):469–472. doi: 10.1016/0005-2744(69)90267-8. [DOI] [PubMed] [Google Scholar]
  4. Colman R. F., Chu R. Deuterium solvent isotope effects in reactions catalyzed. Biochem Biophys Res Commun. 1969 Feb 21;34(4):528–535. doi: 10.1016/0006-291x(69)90414-8. [DOI] [PubMed] [Google Scholar]
  5. Colman R. F. Effect of modification of a methionyl residue on the kinetic and molecular properties of isocitrate dehydrogenase. J Biol Chem. 1968 May 25;243(10):2454–2464. [PubMed] [Google Scholar]
  6. Colman R. F. Evidence for a methionyl residue in the active site of isocitrate dehydrogenase. Biochem Biophys Res Commun. 1967 Jul 21;28(2):222–228. doi: 10.1016/0006-291x(67)90433-0. [DOI] [PubMed] [Google Scholar]
  7. Colman R. F. The role of sulfhydryl groups in the catalytic function of isocitrate dehydrogenase. I. Reaction with 5,5'-dithiobis(2-nitrobenzoic acid). Biochemistry. 1969 Mar;8(3):888–898. doi: 10.1021/bi00831a019. [DOI] [PubMed] [Google Scholar]
  8. DIXON M. GRAPHICAL DETERMINATION OF EQUILIBRIUM CONSTANTS. Biochem J. 1965 Mar;94:760–762. doi: 10.1042/bj0940760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HENDERSON N. S. ISOZYMES OF ISOCITRATE DEHYDROGENASE: SUBUNIT STRUCTURE AND INTRACELLULAR LOCATION. J Exp Zool. 1965 Apr;158:263–273. doi: 10.1002/jez.1401580303. [DOI] [PubMed] [Google Scholar]
  10. Kawahara K. Evaluation of diffusion coefficients of proteins from sedimentation boundary curves. Biochemistry. 1969 Jun;8(6):2551–2557. doi: 10.1021/bi00834a043. [DOI] [PubMed] [Google Scholar]
  11. Magar M. E., Homi M. L. Allosteric inhibition of TPN-linked isocitrate dehydrogenase by folate. Biochem Biophys Res Commun. 1968 Jun 10;31(5):665–670. doi: 10.1016/0006-291x(68)90613-x. [DOI] [PubMed] [Google Scholar]
  12. Magar M. E., Robbins J. E. The subunits of porcine heart TPN-linked isocitrate dehydrogenase. Biochim Biophys Acta. 1969 Sep 30;191(1):173–176. doi: 10.1016/0005-2744(69)90330-1. [DOI] [PubMed] [Google Scholar]
  13. ROSE Z. B. Studies on the mechanism of action of isocitric dehydrogenase. J Biol Chem. 1960 Apr;235:928–933. [PubMed] [Google Scholar]
  14. SIEBERT G., DUBUC J., WARNER R. C., PLAUT G. W. The preparation of isocitric dehydrogenase from mammalian heart. J Biol Chem. 1957 Jun;226(2):965–975. [PubMed] [Google Scholar]

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