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AMP-deaminase from normal and cirrhotic human liver: A comparative study

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Abstract

AMP-deaminase (EC 3.5.4.6) is an enzyme of nucleotide breakdown involved in regulation of adenine nucleotide pool in mammalian cells. Reaction catalysed by AMP-deaminase constitutes a rate-limiting step in adenine nucleotide catabolism in liver. In this study kinetic and regulatory properties of AMP-deaminase purified from normal and cirrhotic human liver were investigated.

In comparison to AMP-deaminase extracted from the normal human liver, AMP-deaminase extracted from the cirrhotic liver was less sensitive towards substrate analogues, and only a very limited response towards pH and adenylate energy charge changes tested for enzyme isolated from this tissue source had been observed. At physiological pH 7.0, in the absence and in the presence of important allosteric effectors (ATP, ADP, GTP and orthophosphate), AMP-deaminases from the two sources studied manifested different regulatory profiles, with half-saturation constant (S0.5) values being distinctly higher for the enzyme extracted from the pathological organ.

In contrast to AMP-deaminase isolated from the normal, healthy liver, where presence of relatively large (68 kDa) protein fragment was also detected, only smaller protein fragments were identified, while SDS-PAG electrophoresis of AMP-deaminase isolated from the cirrhotic liver was performed.

The obtained results indicate clearly that advanced proteolytic processes occurring in the cirrhotic liver may affect structural integrity of AMP-deaminase studied, making enzyme less active and less sensitive to regulatory action of important allosteric effectors. (Mol Cell Biochem 262: 119–126, 2004)

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References

  1. Smith LD, Kizer DE: Purification and properties of rat liver AMP deaminase. Biochim Biophys Acta 191: 415–424, 1969

    Article  PubMed  Google Scholar 

  2. Ronca-Testoni S, Raggi A, Ronca G: Muscle AMP aminohydrolase. III. A comparative study on the regulatory properties of skeletal muscle enzyme from various species. Biochim Biophys Acta 198: 101–112, 1970

    Article  PubMed  Google Scholar 

  3. Yun S, Suelter CH: Human erythrocyte 5'-AMP aminohydrolase. Purification and characterization. J Biol Chem 253: 404–408, 1978

    PubMed  Google Scholar 

  4. Kaletha K, Skladanowski A: Regulatory properties of rat heart AMP deaminase. Biochim Biophys Acta 568: 80–90, 1979

    Article  PubMed  Google Scholar 

  5. Chapman AG, Atkinson DE: Stabilization of the adenylate energy charge by the adenylate deaminase reaction. J Biol Chem 248: 8309–8312, 1973

    PubMed  Google Scholar 

  6. Van den Berghe G, Bronfman M, Vanneste R, Hers H: The mechanism of adenosine triphosphate depletion in the liver after a load of fructose. Biochem J 162: 601–609, 1977

    PubMed  Google Scholar 

  7. Ogasawara N, Goto H, Yamada Y, Watanabe T: Distribution of AMPdeaminase izozymes in rat tissues. Eur J Biochem 87: 297–304, 1978

    PubMed  Google Scholar 

  8. Kaletha K, Nowak G: Developmental forms of human skeletal-muscle AMP deaminase. The kinetic and regulatory properties of the enzyme. Biochem J 249: 255–261, 1987

    Google Scholar 

  9. Marquetant R, Desai NM, Sabina RL, Holmes EW: Evidence for sequential expression of multiple AMP deaminase isoforms during skeletal muscle development. Proc Natl Acad Sci USA 84: 2345–2349, 1987

    PubMed  Google Scholar 

  10. Ogasawara N, Goto H, Yamada Y, Watanabe T, Asano T: AMP deaminase isozymes in human tissues. Biochim Biophys Acta 714: 298–306, 1982

    PubMed  Google Scholar 

  11. Morisaki T, Sabina RL, Holmes EW: Adenylate deaminase. Amultigene family in humans and rats. J Biol Chem 265: 11482–11486, 1990

    PubMed  Google Scholar 

  12. Bausch-Jurken MT, Mahnke-Zizelman DK, Morisaki T, Sabina RL: Molecular cloning of AMP deaminase isoform L. Sequence and bacterial expression of human AMPD2 cDNA. J Biol Chem 267: 22407–22413, 1992

    PubMed  Google Scholar 

  13. Mahnke-Zizelman DK, Sabina RL: Cloning of human AMP deaminase isoform E cDNAs. Evidence for a third AMPD gene exhibiting alternatively spliced 5'-exons. J Biol Chem 267: 20866–20877, 1992

    PubMed  Google Scholar 

  14. Van den Berghe F, Sabina RL: Characterization of human AMP deaminase 2 (AMPD2) gene expression reveals alternative transcripts encoding variable N-terminal extension of isoform L. Biochem J 312: 401–410, 1995

    PubMed  Google Scholar 

  15. Ranieri-Raggi M, Raggi A: Effects of storage on activity and subunit structure of rabbit skeletal muscle AMP deaminase. Biochem J 189: 367–368, 1980

    PubMed  Google Scholar 

  16. Mahnke-Zizelman DK, Tullson PC, Sabina RL: Novel aspects of tetramer assembly and N-terminal domain structure and function are revealed by recombinant expression of human AMP deaminase isoforms. J Biol Chem 273: 35118–35125, 1998

    Article  PubMed  Google Scholar 

  17. Sabina RL, Mahnke-Zizelman DK: Towards an understanding of the functional significance of N-terminal domain divergence in human AMP deaminase isoforms. Pharmacol Ther 87: 279–283, 2000

    Article  PubMed  Google Scholar 

  18. Haas A, Sabina RL: Expression, purification, and inhibition in vitro proteolysis of human AMPD2 (isoform L) recombinant enzymes. Protein Expr Purif 27: 293–303, 2003

    Article  PubMed  Google Scholar 

  19. Szydłowska M, Nagel-Starczynowska G, Rybakowska I, Świeca A, Kaletha K: Human liver AMP-deaminase-oligomeric forms of the enzyme. Mol Cell Biochem 241: 81–86, 2002

    Article  PubMed  Google Scholar 

  20. Smiley KL, Berry AJ, Suelter CH: An improved purification, crystallization, and some properties of rabbit muscle 5'-adenylic acid deaminase. J Biol Chem 242: 2502–2506, 1967

    PubMed  Google Scholar 

  21. Nagel-Starczynowska G, Nowak G, Kaletha K: Purification and properties of AMP-deaminase from human uterine smooth muscle. Biochim Biophys Acta 1073: 470–473, 1991

    Article  PubMed  Google Scholar 

  22. Chaney AL, Marbach EP: Modified reagents for determination of urea and ammonia. Clin Chem 8: 130–132, 1961

    Google Scholar 

  23. Nowak G, Nagel-Starczynowska G, Kaletha K: Purification and properties of AMP-deaminase from human uterine smooth muscle. Regulation by adenylate energy charge and activated fatty acids. Biochim Biophys Acta 1078: 303–304, 1991

    Article  PubMed  Google Scholar 

  24. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976

    Article  CAS  PubMed  Google Scholar 

  25. Szewczyk B, Harper DR: Use of blotted proteins as immunogenes. In: B. Dunbar (ed.). Protein Blotting. A Practical Approach. IRL Press, Oxford, 1994, pp 189–205

    Google Scholar 

  26. Nowak G, Kaletha K: Purification and properties of AMP-deaminase from human kidney. Biochem Med Metab Biol 47: 232–241, 1992

    PubMed  Google Scholar 

  27. Szydłowska M, Chodorowski Z, Rybakowska I, Nagel-Starczynowska G, Kaletha K: Full-size for of human liver AMP-deaminase? Manuscript in preparation

  28. Puig JG, Fox IH: Ethanol-induced activation of adenine nucleotide turnover. evidence for a role of acetate. J Clin Invest 74: 936–941, 1984

    PubMed  Google Scholar 

  29. Puig JG, Mateos FA: Clinical and biochemical aspects of uric acid overproduction. Pharm World Sci 16: 40–54, 1994

    PubMed  Google Scholar 

  30. Rubin E, Farber JL: In: Pathology, 2nd edn. Lippincott, Philadelphia, 1994, p 740

  31. Chapman AG, Miller AL, Atkinson DE: Role of the adenylate deaminase reaction in regulation of adenine nucleotide metabolism in ehrlich ascites tumor. Cells Cancer Res 36: 1144–1150, 1976

    Google Scholar 

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Authors and Affiliations

  1. Department of Anesthesiology and Intensive Therapy, Debinki Medical University of Gdansk, Gdansk, Poland

    Paweł Dutka

  2. Department of Biochemistry Therapy, Debinki Medical University of Gdansk, Gdansk, Poland

    Magdalena Szydłowska

  3. Department of Internal Medicine and Toxicology, Debinki Medical University of Gdansk, Gdansk, Poland

    Zygmunt Chodorowski

  4. Department of Biochemistry Therapy, Debinki Medical University of Gdansk, Gdansk, Poland

    Iwona Rybakowska & Krystian Kaletha

  5. Department of Pharmaceutical Biochemistry, Debinki Medical University of Gdansk, Gdansk, Poland

    Gabriela Nagel-Starczynowska

Authors
  1. Paweł Dutka
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  2. Magdalena Szydłowska
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  3. Zygmunt Chodorowski
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  4. Iwona Rybakowska
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  5. Gabriela Nagel-Starczynowska
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  6. Krystian Kaletha
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Dutka, P., Szydłowska, M., Chodorowski, Z. et al. AMP-deaminase from normal and cirrhotic human liver: A comparative study. Mol Cell Biochem 262, 119–126 (2004). https://doi.org/10.1023/B:MCBI.0000038217.71621.88

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  • DOI: https://doi.org/10.1023/B:MCBI.0000038217.71621.88

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