Lung

, Volume 160, Issue 1, pp 207–218 | Cite as

Acyl CoA synthetase from rat lung purification and properties

  • D. K. Das
Original Articles

Abstract

Long chain fatty acyl-CoA synthetase, a key enzyme in fatty acid metabolism, occurs in liver, brain, muscle, adipose tissue and bacteria. In our study, rat lung was found to have an acyl-CoA synthetase very active for the synthesis of palmitoyl CoA. The lung microsomal and cell membrane fractions were the principle sources of this enzyme, with mitochondria the next most active fraction. Palmitoyl CoA synthetase was isolated from rat lung microsomes and purified 100-fold. Enzyme activity was determined either by spectrophotometric or by radioactive assay methods. Kinetic parameters and properties were determined using purified microsomal palmitoyl CoA synthetase. The assay system required ATP and CoA as substrates. Maximal activation was reached with palmitate as substrate.

Key words

Palmitoyl CoA synthetase Lung Long chain fatty acyl CoA synthetase 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Baginski ES, Foa’ PP, Zak B (1970) In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 876–880Google Scholar
  2. 2.
    Creasey WA (1962) Observations on the activation of stearic acid by rat liver preparations. Biochim Biophys Acta 64:559–561PubMedCrossRefGoogle Scholar
  3. 3.
    Das DK, Ganguly M (1981) Diabetes, hypophysectomy and thyroidectomy reduces L-triiodothyronine binding capacity of rat lung. Endocrinology 109:296–300PubMedCrossRefGoogle Scholar
  4. 4.
    Dole VP (1956) A relation between non-esterified fatty acids in plasma and the metabolism of glucose. J Clin Invest 35:150–159PubMedCrossRefGoogle Scholar
  5. 5.
    Ellman GL (1958) A colorimetric method for determining low concentrations of mercaptans. Arch Biochem Biophys 74:443–450PubMedCrossRefGoogle Scholar
  6. 6.
    Frosolono MF, Slivka S, Charms BL (1971) Acyl transferase activities in dog lung microsomes. J Lipid Res 12:96–103PubMedGoogle Scholar
  7. 7.
    Hogeboom GH (1955) In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 1. Academic Press, New York, p 16CrossRefGoogle Scholar
  8. 8.
    Kornberg A, Pricer WE Jr (1953) Enzymatic synthesis of the coenzyme A derivatives of long chain fatty acids. J Biol Chem 204:329–343PubMedGoogle Scholar
  9. 9.
    Kumar S, Das DK, Dorfman AE, Asato N (1977) Stimulation of the synthesis of hepatic fatty acid synthesizing enzymes of hypophysectomized rats by 3,5,3′-L-triiodothyronine. Arch Biochem Biophys 178:507–516PubMedCrossRefGoogle Scholar
  10. 10.
    Lands WFM (1960) Metabolism of glycerolipids 11. The enzymatic acylation of lysolecithin. J Biol Chem 235:2233–2237PubMedGoogle Scholar
  11. 11.
    Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666CrossRefGoogle Scholar
  12. 12.
    Lippel K, Robinson J, Trams EG (1970) Intracellular distribution of palmitoyl CoA synthetase in rat liver. Biochim Biophys Acta 206:173–177PubMedGoogle Scholar
  13. 13.
    Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin Phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  14. 14.
    McCaman RE, McCaman MW, Hunt JM, Smith MS (1965) Microdetermination of monoamine oxidase and 5-hydroxytryptophan decarboxylase activities in nervous tissue. J Neurochem 12:15–19PubMedCrossRefGoogle Scholar
  15. 15.
    Oldenborg V, Van Golde LMG (1976) Activity of choline phosphotransferase, lysolecithin lysolecithin acyltransferase and lysolecithin acyltransferase in the developing mouse lung. Biochim Biophys Acta 441:433–442PubMedGoogle Scholar
  16. 16.
    Ouchterlony O (1949) Antigen-antibody reactions in gels. Acta Pathol Microbiol Scand 26:507–515PubMedGoogle Scholar
  17. 17.
    Pande SV, Mead JF (1968) Distribution of long-chain fatty acid-activating enzymes in rat tissues. Biochim Biophys Acta 152:636–638PubMedGoogle Scholar
  18. 18.
    Pande SV, Mead JF (1968) Long chain fatty acid activation in subcellular preparations from rat liver. J Biol Chem 243:352–361PubMedGoogle Scholar
  19. 19.
    Samuel D, Estroumza J, Ailhaud G (1970) Partial purification and properties of Acyl CoA synthetase ofEscherichia coli. Eur J Biochem 12:576–582PubMedCrossRefGoogle Scholar
  20. 20.
    Van Golde LMG (1976) Metabolism of phospholipids in the lung. Am Rev Respir Dis 114:977–1000PubMedGoogle Scholar
  21. 21.
    Vignais PV, Gallagher CH, Zabin I (1958) Activation and oxidation of long chain fatty acids by rat brain. J Neurochem 2:283–287PubMedCrossRefGoogle Scholar
  22. 22.
    Vignais PV, Zabin I (1958) Synthesis and properties of palmityl adenylate, palmityl coenzyme A and palmityl glutathione. Biochim Biophys Acta 29:263–269PubMedCrossRefGoogle Scholar
  23. 23.
    Weiss SB, Kennedy EP, Kiyasu JY (1960) The enzymatic synthesis of triglycerides. J Biol Chem 235:40–44PubMedGoogle Scholar
  24. 24.
    Yates DW, Shepherd D, Garland PB (1966) Organization of fatty acid activation in rat liver mitochondria. Nature 209:1213–1215PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • D. K. Das
    • 1
  1. 1.Division of Pulmonary MedicineLong Island Jewish-Hillside Medical Center and State University of New York at Stony BrookNew Hyde ParkUSA

Personalised recommendations