Abstract
AMP-degrading pathways in Azotobacter vinelandii cells were investigated. AMP nucleosidase (EC 3.2.2.4) was rapidly synthesized and reached a maximum at 24 h, while the activity of 5′-nucleotidase (EC 3.1.3.5) specific for AMP, which was negligible during the logarithmic phase of the growth, first appeared in 24 h-cultures, and reached a maximum after complete exhaustion of sucrose from the growth medium (70 h).
Cell-free extracts of A. vinelandii of 48 h-cultures hydrolyzed AMP to ribose 5-phosphate and adenine in the presence of ATP, and adenine was deaminated to hypoxanthine. When ATP was excluded, AMP was dephosphorylated to adenosine, which was further metabolized to inosine, and finally to hypoxanthine. Hypoxanthine thus formed was reutilized for the salvage synthesis of IMP under the conditions where 5-phosphoribosyl 1-pyrophosphate was able to be supplied. These results suggest that the levels of ATP can determine the rate of AMP degradation by the AMP nucleosidase- and 5′-nucleotidase-pathways. The role of ATP in the AMP degradation was discussed in relation to the regulatory properties of AMP nucleosidase, inosine nucleosidase (EC 3.2.2.2) and adenosine deaminase (EC 3.5.4.4).
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References
Ashwell, G.: Colorimetric analysis of sugars. In: Methods in enzymology, Vol. 3, (Colowick, S. P. and Kaplan, N. O., eds.), pp. 73–105. New York: Academic Press 1957
Atkinson, D. E.: Cellular energy metabolism and its regulation. New York: Academic Press 1977
Chaney, A. L., Marbach, E. P.: Modified reagents for determination of urea and ammonia. Clin. Chem. 8, 130–132 (1962)
Chapman, A. G., Atkinson, D. E.: Stabilization of adenylate energy charge by the adenylate deaminase reaction. J. Biol. Chem. 248, 8309–8312 (1973)
Chen, P. S., Jr., Toribara, T. Y.: Microdetermination of phosphorus. Anal. Chem. 28, 1756–1758 (1956)
Cowsert, M. K., Jr., Carrier, O., Jr., Crowell, J. W.: The effect of hemorrhagic shock on blood uric acid level. Canad. J. Physiol. Pharmacol. 44, 861–864 (1966)
Crowell, J. W., Jones, C. E., Smith, E. E.: Effect of allopurinol on hemorrhagic shock. Am. J. Physiol. 216, 744–748 (1969)
Dawson, R. M. C., Elliot, D. C., Elliot, W. H., Jones, K. M. (eds.): Data for biochemical research. Oxford: Clarendon Press 1969
Dygert, S., Li, L. H., Florida, D., Thoma, J. A.: Determination of reducing sugar with improved precision. Anal. Biochem. 13, 367–374 (1965)
Frick, G. P., Lowentein, J. M.: Vectoral production of adenosine by 5′-nucleotidase in perfused rat heart. J. Biol. Chem. 253, 1240–1244 (1978)
Heppel, L. A., Hurwitz, J., Horecker, B. L.: Adenine deaminase of Azotobacter vinelandii. J. Am. Chem. Soc. 79, 630–633 (1957)
Kohn, M. C., Garfinkel, D.: Computer simulation of ischemic rat heart metabolism. I. Model construction. Am. J. Physiol. 232, 386–393 (1977a)
Kohn, M. C., Garfinkel, D.: Computer simulation of ischemic rat heart metabolism. II. Model behavior. Am. J. Physiol. 232, 394–399 (1977b)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with Folin-phenol reagent. J. Biol. Chem. 193, 265–272 (1951)
Murakami, K., Yoshino, M.: Ion-dependent activation of AMP nucleosidase from Azotobacter vinelandii. Biochim. Biophys. Acta 613, 153–159 (1980)
Schramm, V. L., Leung, H.: Regulation of adenosine monophosphate levels as a function of adenosine triphosphate and inorganic phosphate. A proposed metabolic role of adenosine monophosphate nucleosidase from Azotobacter vinelandii. J. Biol. Chem. 248, 8313–8315 (1973)
Schramm, V. L., Lazorik, F. C.: The pathway of adenylate catabolism in Azotobacter vinelandii. Evidence for adenosine monophosphate nucleosidase as the regulating enzyme. J. Biol. Chem. 250, 1801–1808 (1975)
Swissa, M., Weinhouse, H., Benziman, M.: Activities of citrate synthase and other enzymes of Acetobacter xylinum in situ and in vitro. Biochem. J., 153, 499–501 (1976)
Tsukada, T., Yoshino, M.: Adenosone deaminase form Azotobacter vinelandii. Purification and Properties. Arch. Microbiol. 128, 228–232 (1980)
Woods, H. F., Eggleston, L. K., Krebs, H. A.: The cause of hepatic accumulation of fructose 1-phosphate in fructose loading. Biochem. J. 119, 501–510 (1970)
Yoshino, M., Ogasawara, N., Suzuki, N., Kotake, Y.: Regulation of AMP nucleosidase in Azotobacter vinelandii. Biochim. Biophys. Acta 146, 620–622 (1967)
Yoshino, M.: AMP nucleosidase from Azotobacter vinelandii. I. Purification and properties. J. Biochem. 68, 321–329 (1970)
Yoshino, M., Ogasawara, N.: AMP nucleosidase from Azotobacter vinelandii. III. Kinetics of allosteric interactions. J. Biochem. 72, 223–233 (1972)
Yoshino, M., Kawamura, Y., Ogasawara, N.: Regulation of AMP deaminase from chicken erythrocytes. A kinetic study of the allosteric interactions. J. Biochem. 80, 299–308 (1976)
Yoshino, M., Murakami, K., Tsushima, K.: The role of polyamines in the regulation of AMP deaminase isozymes. Biochim. Biophys. Acta 542, 177–179 (1978a)
Yoshino, M., Tsukada, T., Tsushima, K.: Inosine nucleosidase from Azotobacter vinelandii. Purification and properties. Arch. Microbiol. 119, 59–64 (1978b)
Yoshino, M., Murakami, K., Tsushima, K.: Polyamines as activators of AMP nucleosidase from Azotobacter vinelandii. Experientia 35, 578–579 (1979a)
Yoshino, M., Murakami, K., Tsushima, K.: Effect of monovalent cations on AMP nucleosidase from Azotobacter vinelandii. Biochim. Biophys. Acta 570, 118–123 (1979b)
Yoshino, M., Murakami, K., Tsushima, K.: AMP deaminase from baker's yeast. Purification and some regulatory properties. Biochim. Biophys. Acta 570, 157–166 (1979c)
Zielke, C. L., Suelter, C. H.: Purine purine nucleoside, and purine nucleotide aminohydrolases. In: The Enzymes, Vol. 4, 3rd edn. (Boyer, P. D., ed.), pp. 47–78. New York: Academic Press 1971
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Yoshino, M., Tsukada, T., Murakami, K. et al. Adenine nucleotide metabolism in Azotobacter vinelandii. Two metabolic pathways of AMP degradation. Arch. Microbiol. 128, 222–227 (1980). https://doi.org/10.1007/BF00406162
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DOI: https://doi.org/10.1007/BF00406162