Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Isolation, Purification, and Partial Characterization of an AMP Deaminase from Saccharomyces cerevisiae D

  • Kim, Myung-Hee (Microbial Enzyme Research Unit, Korea Research Institute of Bioscience & Biotechnology) ;
  • Lee, Jung-Kee (Microbial Enzyme Research Unit, Korea Research Institute of Bioscience & Biotechnology) ;
  • Kim, Hyung-Kwoun (Microbial Enzyme Research Unit, Korea Research Institute of Bioscience & Biotechnology) ;
  • Oh, Tae-Kwang (Microbial Enzyme Research Unit, Korea Research Institute of Bioscience & Biotechnology)
  • Published : 1999.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

An adenosine 5'-monophosphate deaminase (AMP aminohydrolase, EC 3.5.4.6) was purified to homogeneity from the cell-free extract of Saccharomyces cerevisiae DKCTC7248. The molecular mass of subunit was estimated to be 80 kDa on SDS-PAGE, and that of the holoenzyme was shown to be 240 kDa by gel filtration. The isoelectric point of the enzyme (AMP deaminase D) was determined to be 6.2. The AMP deaminase D was specific towards AMP with an apparent $K_m$ value of 4.1 mM and a Hill coefficient, $n_H$, of 2.2. Both ATP and ADP were positive allosteric effectors of the AMP deaminase D: The apparent $K_{m}$ was decreased to 1.6 mM and 3.3 mM in the presence of 0.1 mM ATP and ADP, respectively, lowering $n_{H}$ to 1.0. Univalent cations like $K^+, Na^+ and Li^ +$ activated the enzyme but some divalent cations such as $Cu^{ 2+} and Cd^{2+}$ showed strong inhibitory effects. This enzyme displayed optimum activity at $30^{\circ}C$ and pH 7.0. In addition, it was stable up to $45^{\circ}C$ and over a wide pH range(pH 5.5-9.0). Amino acid sequences of its N-terminal region were analyzed to be ADYKMQMFADDA.

Keywords

References

  1. J. Allergy Clin. Immunol v.80 Monosidium-induced asthma Allen, D. H.;J. Delohery;G. Baker
  2. Appl. Environ.Microbiol v.62 Evaluation of molecular typing techniques to assign genetic diversity among Saccharomyce cerevisiae strains Baleiras couto,M. M.;B. Eijsma;H. Hofstra;J. H. J. Huis in't Veld;J. M. B. M. van der Vossen
  3. Anal. Biochem v.72 A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye staining Bradford, M. M.
  4. J. Bacteriol v.155 Purine salvage pathways of Bacillus subtilis and effect of guanine on growth of GMP-reductase mutants Endo, T.;B. Uratani;E. Freese
  5. J. Comp. Physiol v.163 Comparative enzymology of AMP deaminase, adenylate kinase, and creatine kinase in vertebrate heart and skeletal muscle: the characteristic AMP deaminase levels of skeletal versus cardiac muscle are reversed in the North American toad Fishbein, W. N.;J. I. Davis;J. W. Foellmer
  6. J. Microbiol. Biotechnol v.8 Invertase production by fed-batch fermentations of recombinant Saccharomyces cerevisiae Koo, J. H.;S. Y. Kim;Y. C. Park;N. S. Han;J. H. Seo
  7. Nature v.227 Cleavage of structural protein during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  8. J. Biol. Chem v.264 AMP deaminase from yeast Mcrkler, D. J.;A. S. Walj;J. Taylor;V. L. Schramm
  9. Biochemistry v.28 Characterization of AMD, the AMP deaminase gene in yeast. Production of amd strain, cloning, nucleotide sequence, and properties of the protein Meyer, S. L.;K. L. Kvalnes-Krick;V. L. Schramm
  10. Biochem. Biophys. Acta v.1073 Purification and properties of AMP deaminase from human uterine smooth muscle Nagel-Starczynowska, G.;G. Nowak;K. Kaletha
  11. Biochem, Biophys. Acta v.1164 AMP-deaminase form human uterine smooth muscle:The effect of DTNB treatment on kinetic and regulatory properties of the enzyme Nagel-Starczynowska, G.;K. Kaletha
  12. Eur. J. Biochem v.87 Distribution of AMP-deaminase isozymes in rat tissues Ogasawara, N.;H. Goto;Y. Yamada;T. Watanabe
  13. World J. Microbiol. Biotechnol v.10 Contimuius production of 5'-ribonucleotides from teast RNA by hydrolysis with immobilized 5'-phosphodiesterase and 5'-adenylate deaminase Olemedo, F.;F. Iturbe;J. Gomez-Hernandez;A. Lopez-Munguia
  14. Comp. Biochem. Physiol v.99B A specific AMP deaminase assay and its application to tissue homogenates Raffin, J. P.;M. T. Thebault
  15. Biochem. J. v.309 Regulation of skeletal-muscle AMP deaminase: involvement of histidine residues in the pH-dependent inhibition of the rabbit enzyme by ATP Ranieri-Raggi, M.;F. Ronca;A. Sabbatini;A. Raggi
  16. Protein Structure, A Practical Approach Righetti, P. G.;T. E. Creighton(ed.)
  17. J. Biol. Chem v.251 Purification of human erythrocyte adenosine deaminase by affinity column chromatography Schrader, W. P.;A. R. Stacy;B. Pollara
  18. J. Biol. Chem v.242 Adenylate deaminase Ⅱ. Purification and some regulatory properties of the enzyme from calf brain Setlow, B.;J. Lowenstein
  19. Neurokhimiya v.11 Isolation of AMP deaminase molecular forms from the cattle nerve tossue Sharoyan, S.;S. Mardanian;A. Haroutunian
  20. Biochem. J. v.290 Isolation and characterization of AMP deaminase from mammalian (rabbit) myocardium Thakkar, J. K.;D. R. Janero;C. Yarwood;H. sharif;D. Hreniuk
  21. Acta Neuropathol. Berlin v.75 Neurotoxicity of monosodium-L-glutamate in pregnant and fetal rats Toth, L.;S. Karcsu;J. Feledi;G. W. Kreutzberg
  22. Appl. Environ. Microbiol v.61 Genetic diversity and geographical distribution of wild Saccharomyces cerevisiae strains from the wine-producing area of Charentes Versavaud, A.;P. Courcoux;C. Roulland;L. Dulau;J. N. Hallet
  23. Phytochemistry v.31 AMP deaminase and the control of adenylate catabolism in suspension-cultured Catharathus roseus cells Yabuki, N.;H. Ashihra
  24. Biochim. Biophys. Acta v.570 AMP deaminase from baker's yeast; purification and some regulatory properties Yoshino, M.;K. Murakami;K. Tsushima