Partial Purification and Properties of an AMP-specific Soluble 5'-nucleotidase from Pigeon Heart

Overview

Journal Biochem J

Specialty Biochemistry

Date 1990 May 15

PMID 2344353

Citations 12

Authors

A C Skladanowski

A C Newby

Affiliations

Soon will be listed here.

Abstract

A soluble 5'-nucleotidase was purified 200-fold from pigeon heart. The enzyme (1) had an apparent molecular mass close to 150 kDa, (2) had a neutral pH optimum and hydrolysed a wide range of nucleoside 5'-monophosphates with a 15-fold preference for AMP over IMP, (3) at near-physiological concentrations of AMP was activated by ADP but not by ATP, (4) was inhibited by high Mg2+ concentration and high ionic strength, (5) was weakly inhibited by p-nitrophenol phosphate and Pi, and (6) was non-competitively inhibited more potently by 5'-deoxy-5'-isobutylthioinosine than by 5'-deoxy-5'-isobutylthioadenosine, but not by [alpha,beta-methylene]ADP. The data show that the enzyme is distinct from the ecto-5'-nucleotidase and from the previously purified IMP-specific 5'-nucleotidase. They also predict that the enzyme is activated during ATP catabolism and hence will generate a more-than-linear increase in the adenosine-formation rate in response to an increase in cytosolic AMP concentration.

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Structure-activity relationship of cytoplasmic 5'-nucleotidase substrate sites.

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Effect of 5'-deoxy-5'-isobutylthioadenosine on formation and release of adenosine from neonatal and adult rat ventricular myocytes.

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References

Imai S, Chin W, Jin H, Nakazawa M . Production of AMP and adenosine in the interstitial fluid compartment of the isolated perfused normoxic guinea pig heart. Pflugers Arch. 1989; 414(4):443-9. DOI: 10.1007/BF00585055. View

Headrick J, Willis R . 5'-Nucleotidase activity and adenosine formation in stimulated, hypoxic and underperfused rat heart. Biochem J. 1989; 261(2):541-50. PMC: 1138859. DOI: 10.1042/bj2610541. View

Stochaj U, Flocke K, Mathes W, Mannherz H . 5'-Nucleotidases of chicken gizzard and human pancreatic adenocarcinoma cells are anchored to the plasma membrane via a phosphatidylinositol-glycan. Biochem J. 1989; 262(1):33-40. PMC: 1133225. DOI: 10.1042/bj2620033. View

Dixon M . The determination of enzyme inhibitor constants. Biochem J. 1953; 55(1):170-1. PMC: 1269152. DOI: 10.1042/bj0550170. View

Itoh R . Regulation of cytosol 5'-nucleotidase by adenylate energy charge. Biochim Biophys Acta. 1981; 659(1):31-7. DOI: 10.1016/0005-2744(81)90268-0. View

Itoh R . Studies on some molecular properties of cytosol 5'-nucleotidase from rat liver. Biochim Biophys Acta. 1982; 716(1):110-3. DOI: 10.1016/0304-4165(82)90208-2. View

Worku Y, Newby A . Nucleoside exchange catalysed by the cytoplasmic 5'-nucleotidase. Biochem J. 1982; 205(3):503-10. PMC: 1158514. DOI: 10.1042/bj2050503. View

Worku Y, Newby A . The mechanism of adenosine production in rat polymorphonuclear leucocytes. Biochem J. 1983; 214(2):325-30. PMC: 1152251. DOI: 10.1042/bj2140325. View

Itoh R, Oka J . Evidence for existence of a cytosol 5'-nucleotidase in chicken heart: comparison of some properties of heart and liver enzymes. Comp Biochem Physiol B. 1985; 81(1):159-63. DOI: 10.1016/0305-0491(85)90177-4. View

10.

Meghji P, Holmquist C, Newby A . Adenosine formation and release from neonatal-rat heart cells in culture. Biochem J. 1985; 229(3):799-805. PMC: 1145127. DOI: 10.1042/bj2290799. View

11.

Madrid-Marina V, Fox I . Human placental cytoplasmic 5'-nucleotidase. Kinetic properties and inhibition. J Biol Chem. 1986; 261(1):444-52. View

12.

Bunger R, Soboll S . Cytosolic adenylates and adenosine release in perfused working heart. Comparison of whole tissue with cytosolic non-aqueous fractionation analyses. Eur J Biochem. 1986; 159(1):203-13. DOI: 10.1111/j.1432-1033.1986.tb09854.x. View

13.

Itoh R, Oka J, Ozasa H . Regulation of rat heart cytosol 5'-nucleotidase by adenylate energy charge. Biochem J. 1986; 235(3):847-51. PMC: 1146764. DOI: 10.1042/bj2350847. View

14.

FRITZSON P, Haugen T, Tjernshaugen H . The presence and activity in normal and regenerating rat liver postmicrosomal supernatant fraction of an enzyme with properties similar to those of membrane-bound 5'-nucleotidase. Biochem J. 1986; 239(1):185-90. PMC: 1147258. DOI: 10.1042/bj2390185. View

15.

Le Hir M, Dubach U . An ATP-inhibited soluble 5'-nucleotidase of rat kidney. Am J Physiol. 1988; 254(2 Pt 2):F191-5. DOI: 10.1152/ajprenal.1988.254.2.F191. View

16.

Meghji P, Middleton K, Newby A . Absolute rates of adenosine formation during ischaemia in rat and pigeon hearts. Biochem J. 1988; 249(3):695-703. PMC: 1148762. DOI: 10.1042/bj2490695. View

17.

Itaya K, Ui M . A new micromethod for the colorimetric determination of inorganic phosphate. Clin Chim Acta. 1966; 14(3):361-6. DOI: 10.1016/0009-8981(66)90114-8. View

18.

GIBSON W, DRUMMOND G . Properties of 5'-nucleotidase from avian heart. Biochemistry. 1972; 11(2):223-9. DOI: 10.1021/bi00752a013. View

19.

Burger R, Lowenstein J . 5'-Nucleotidase from smooth muscle of small intestine and from brain. Inhibition of nucleotides. Biochemistry. 1975; 14(11):2362-6. DOI: 10.1021/bi00682a014. View

20.

Naito Y, Tsushima K . Cytosol 5'-nucleotidase from chicken liver. Purification and some properties. Biochim Biophys Acta. 1976; 438(1):159-68. DOI: 10.1016/0005-2744(76)90232-1. View