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Yan LJ, Thangthaeng N, Sumien N, et al. Serum Dihydrolipoamide Dehydrogenase Is a Labile Enzyme. J Biochem Pharmacol Res. 2013;1(1):30-42
Yan, L. J., Thangthaeng, N., Sumien, N., & Forster, M. J. (2013). Serum Dihydrolipoamide Dehydrogenase Is a Labile Enzyme. Journal of biochemical and pharmacological research, 1(1), 30-42.
Yan, Liang-Jun, et al. "Serum Dihydrolipoamide Dehydrogenase Is a Labile Enzyme." Journal of biochemical and pharmacological research vol. 1,1 (2013): 30-42.
Yan LJ, Thangthaeng N, Sumien N, Forster MJ. Serum Dihydrolipoamide Dehydrogenase Is a Labile Enzyme. J Biochem Pharmacol Res. 2013 Mar;1(1):30-42. PMID: 23646291; PMCID: PMC3641859.
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J Biochem Pharmacol Res. 2013 Mar;1(1):30-42.

Serum Dihydrolipoamide Dehydrogenase Is a Labile Enzyme.

Journal of biochemical and pharmacological research

Liang-Jun Yan, Nopporn Thangthaeng, Nathalie Sumien, Michael J Forster

Affiliations

  1. Department of Pharmacology and Neuroscience and Institute for Aging and Alzheimer's Disease Research, University of North Texas Health Science Center, Fort Worth, Texas, USA.

PMID: 23646291 PMCID: PMC3641859

Abstract

Dihydrolipoamide dehydrogenase (DLDH) is a multifunctional oxidoreductase and is well known as an essential component of four mammalian mitochondrial multienzyme complexes: pyruvate dehydrogenase, α-ketoglutarate dehydrogenase, branched chain α-keto acid dehydrogenase, and the glycine cleavage system. However, existence of extracellular DLDH in mammals, if any, has not been clearly defined. The present article reports identification and biochemical characterization of serum DLDH. Proteomic analysis of rat serum using blue native polyacrylamide gel electrophoresis (BN-PAGE) and mass spectrometry peptide sequencing led to generation of 6 tryptic peptides in one band that matched to mitochondrial DLDH, indicating the existence of DLDH in rat serum. Measurement of enzymatic activity also indicated the existence of DLDH in human and mouse serum. Further biochemical analysis of rat serum DLDH revealed that this enzyme lacked diaphorase activity and could not be detected on Western blots probed with antibodies that recognized mitochondrial DLDH. Moreover, both ammonium sulfate fractioning and gel filtration of serum samples rendered a great loss in DLDH activity, indicating that the enzyme activity of this serum protein, unlike that of mitochondrial DLDH, is very labile. When DTT was supplemented in the buffer used for gel filtration, DLDH activity was found to be largely preserved; indicating that serum DLDH is susceptible to air-implicated inactivation. Results of the present study indicate that serum DLDH differs from mitochondrial DLDH in that it is a very labile enzyme.

Keywords: blue native polyacrylamide gel electrophoresis; dihydrolipoamide dehydrogenase; mitochondria; serum

References

  1. Eur J Biochem. 1972 Jul 13;28(2):253-60 - PubMed
  2. J Bacteriol. 1989 Mar;171(3):1346-54 - PubMed
  3. J Neurosci Res. 2007 Aug 1;85(10):2216-23 - PubMed
  4. Antioxid Redox Signal. 1999 Fall;1(3):371-9 - PubMed
  5. Biochem J. 1989 Feb 1;257(3):625-32 - PubMed
  6. Arch Biochem Biophys. 1992 Dec;299(2):356-60 - PubMed
  7. PLoS One. 2008 May 14;3(5):e2161 - PubMed
  8. Biochim Biophys Acta. 1971 Feb 10;227(2):248-63 - PubMed
  9. J Biol Chem. 1969 Aug 25;244(16):4351-9 - PubMed
  10. J Biol Chem. 2005 Jul 8;280(27):25743-53 - PubMed
  11. Biochem Biophys Res Commun. 1986 Mar 28;135(3):802-7 - PubMed
  12. Clin Chem. 1976 Feb;22(2):275-7 - PubMed
  13. Free Radic Biol Med. 2001 May 1;30(9):992-9 - PubMed
  14. Clin Chem. 1984 Nov;30(11):1817-20 - PubMed
  15. J Biol Chem. 1969 May 25;244(10):2511-5 - PubMed
  16. J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Feb 15;879(5-6):386-94 - PubMed
  17. Sheng Wu Wu Li Hsueh Bao. 2012 Apr 20;28(4):341-350 - PubMed
  18. J Bacteriol. 1981 Nov;148(2):639-46 - PubMed
  19. Free Radic Biol Med. 2009 Oct 15;47(8):1147-53 - PubMed
  20. J Biochem. 1991 Mar;109(3):450-4 - PubMed
  21. Electrophoresis. 2007 Apr;28(7):1036-45 - PubMed
  22. Biol Pharm Bull. 1999 Aug;22(8):883-5 - PubMed
  23. Mol Microbiol. 2005 Jan;55(1):27-38 - PubMed
  24. FEMS Microbiol Lett. 2008 Apr;281(2):147-54 - PubMed
  25. FEBS Lett. 1999 Apr 1;448(1):190-2 - PubMed
  26. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
  27. Biochim Biophys Acta. 1972 May 25;267(2):249-57 - PubMed
  28. Proteomics. 2011 Dec;11(23):4468-76 - PubMed
  29. Chem Biol Interact. 2002 Aug 15;140(3):199-213 - PubMed
  30. Mech Ageing Dev. 2008 May;129(5):282-90 - PubMed
  31. Biochim Biophys Acta. 2012 Oct;1817(10):1879-85 - PubMed
  32. J Neurol Neurosurg Psychiatry. 1981 Jul;44(7):574-82 - PubMed
  33. Protein Expr Purif. 2009 Oct;67(2):132-8 - PubMed
  34. Clin Chim Acta. 1969 Jul;25(1):11-6 - PubMed
  35. Arch Biochem Biophys. 1989 Feb 1;268(2):409-25 - PubMed
  36. Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):12896-901 - PubMed
  37. Biochemistry. 1994 Mar 1;33(8):2240-6 - PubMed
  38. FEBS J. 2011 Nov;278(22):4243-51 - PubMed
  39. FEBS J. 2008 Jul;275(13):3278-89 - PubMed
  40. Am J Physiol Regul Integr Comp Physiol. 2004 Nov;287(5):R1244-9 - PubMed
  41. Biochem J. 1987 May 1;243(3):661-5 - PubMed
  42. Biochemistry. 1968 Apr;7(4):1322-7 - PubMed
  43. Biochim Biophys Acta. 1966 Aug 10;122(2):355-8 - PubMed
  44. Biochem Pharmacol. 1992 Aug 18;44(4):631-5 - PubMed
  45. Biochemistry. 2003 Feb 25;42(7):2218-28 - PubMed
  46. Can J Neurol Sci. 1980 Nov;7(4):413-6 - PubMed
  47. Hum Mol Genet. 2011 Aug 1;20(15):2984-95 - PubMed
  48. J Neurochem. 2004 Mar;88(6):1352-60 - PubMed
  49. Structure. 2006 Jan;14(1):129-39 - PubMed
  50. Biochemistry. 1998 Mar 24;37(12):4224-34 - PubMed
  51. Free Radic Res. 2011 Jan;45(1):53-8 - PubMed
  52. Am J Respir Cell Mol Biol. 2001 May;24(5):627-32 - PubMed
  53. Plant Physiol. 2001 Oct;127(2):615-23 - PubMed
  54. Free Radic Res. 2013 Feb;47(2):123-33 - PubMed
  55. Aging Cell. 2013 Feb;12(1):130-8 - PubMed
  56. Anal Biochem. 1998 Oct 1;263(1):67-71 - PubMed
  57. Methods Enzymol. 1995;252:186-95 - PubMed
  58. J Biochem. 1996 Jun;119(6):1118-23 - PubMed
  59. J Biol Chem. 1973 Jul 10;248(13):4834-9 - PubMed
  60. Arch Biochem Biophys. 1987 Aug 1;256(2):597-605 - PubMed
  61. J Biochem. 1982 Feb;91(2):553-61 - PubMed
  62. Biochem Cell Biol. 1992 Jan;70(1):70-5 - PubMed
  63. FEBS Lett. 2012 Mar 9;586(5):585-95 - PubMed
  64. FEBS J. 2011 Nov;278(22):4230-42 - PubMed
  65. Free Radic Biol Med. 1997;22(3):535-42 - PubMed
  66. Biochim Biophys Acta. 1979 Aug 14;547(2):411-6 - PubMed
  67. FEBS Lett. 2004 Jun 18;568(1-3):146-50 - PubMed
  68. Proc Natl Acad Sci U S A. 2007 Apr 10;104(15):6158-63 - PubMed
  69. Plant Physiol. 2010 Feb;152(2):747-61 - PubMed
  70. Science. 2002 Feb 8;295(5557):1073-7 - PubMed
  71. Eur J Biochem. 2001 Mar;268(5):1486-90 - PubMed
  72. J Cardiovasc Pharmacol. 2009 Nov;54(5):391-8 - PubMed
  73. J Mol Recognit. 2009 Mar-Apr;22(2):138-45 - PubMed
  74. Eur J Biochem. 1986 Apr 15;156(2):399-405 - PubMed
  75. J Bacteriol. 1991 May;173(10):3109-16 - PubMed
  76. Biochim Biophys Acta. 2013 Feb;1833(2):253-9 - PubMed
  77. J Bacteriol. 1989 Feb;171(2):665-8 - PubMed
  78. Indian J Biochem Biophys. 1996 Jun;33(3):168-76 - PubMed
  79. J Biol Chem. 2002 Mar 22;277(12):10064-72 - PubMed
  80. J Pharmacol Exp Ther. 2003 Oct;307(1):67-73 - PubMed
  81. Neurology. 1981 Jun;31(6):783-5 - PubMed
  82. Biochim Biophys Acta. 2011 Mar;1808(3):1012-20 - PubMed
  83. J Biol Chem. 1967 Jan 10;242(1):54-60 - PubMed
  84. Anal Biochem. 1985 Oct;150(1):76-85 - PubMed
  85. Biochem Soc Trans. 1988 Apr;16(2):87-9 - PubMed
  86. Biochim Biophys Acta. 2007 Sep;1767(9):1126-33 - PubMed
  87. Front Plant Sci. 2012 Sep 25;3:221 - PubMed
  88. J Biol Chem. 1965 Dec;240(12):4793-800 - PubMed
  89. Eur J Biochem. 1983 Oct 3;135(3):519-27 - PubMed
  90. J Biochem Pharmacol Res. 2013 Mar;1(1):15-26 - PubMed
  91. Gen Physiol Biophys. 2009 Dec;28(4):384-90 - PubMed
  92. Mult Scler. 2010 Apr;16(4):387-97 - PubMed
  93. Biochem J. 1960 Nov;77:341-51 - PubMed
  94. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 - PubMed
  95. FEBS Lett. 2011 Jan 21;585(2):385-9 - PubMed
  96. Anal Biochem. 2009 Jun 15;389(2):143-9 - PubMed
  97. Eur J Biochem. 1989 Jan 15;179(1):61-9 - PubMed
  98. J Biol Chem. 1991 May 25;266(15):9367-73 - PubMed
  99. J Biol Chem. 2011 Nov 18;286(46):40232-45 - PubMed
  100. Biochem Cell Biol. 1992 Aug;70(8):656-63 - PubMed
  101. Science. 2012 Nov 9;338(6108):807-10 - PubMed
  102. J Biol Chem. 2005 Apr 22;280(16):16106-14 - PubMed

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