Skip to main content
SpringerLink
Log in

Partial purification and properties of long-chain acyl-CoA hydrolase from rat brain cytosol

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Long-chain acyl-CoA hydrolase (EC 3.1.2.2.) has been partially purified from the 100,000 × g supernatant fraction of rat brain tissue. The purification procedure included chromatography on gel filtration media, DEAE-cellulose, CM-cellulose, and hydroxyapatite. The partially purified enzyme had a specific activity of 7.1 μmol/min-mg, and when analyzed by polyacrylamide gel electrophoresis, revealed one major and three minor bands of protein in the presence of dodecyl sulfate and two major bands of protein in the absence of dodecyl sulfate. The enzyme had a molecular weight of 65,000 and showed no evidence of aggregated or dissociated forms. The highest catalytic activity was exhibited with palmitoyl-CoA and oleoyl-CoA as substrates. Lower activity was found with decanoyl-CoA as the substrate and little or no activity was found with acetyl-CoA, malonyl-CoA, butyryl-CoA, or acetoacetyl-CoA. The enzyme was inhibited by CoA, various metal ions, including Mn2+, Mg2+ and Ca2+, and by bovine serum albumin. Heating the enzyme produced a loss of activity which corresponded to a first-order kinetic process, the rate of which was independent of the choice of substrate used to measure enzyme activity. This finding supports the idea that the purification procedure yields a single species of long-chain acyl-CoA hydrolase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Kurooka, S., Hosoki, K., andYoshimura, Y. 1972. Some Properties of Long Chain Fatty Acyl-Coenzyme A Thioesterase in Rat Organs. J. Biochem. (Tokyo) 71:625–634.

    Google Scholar 

  2. Colli, W., Hinkle, P. C., andPullman, M. E. 1969. Characterization of the Fatty Acid Elongation System in Soluble Extracts and Membrane Preparations of Rat Liver Mitochondria. J. Biol. Chem. 214:6432–6443.

    Google Scholar 

  3. Mahadevan, E., andSauer, F. 1969. Carnitine Ester Hydrolase of Rat Liver. J. Biol. Chem. 244:4448–4453.

    Google Scholar 

  4. Berge, R. K., andFarstad, M. 1979. Dual Localization of Long-Chain Acyl-CoA Hydrolase in Rat Liver: One in the Microsomes and One in the Mitochondrial Matrix. Eur. J. Biochem. 95:89–97.

    Google Scholar 

  5. Berge, R. K., Slinde, F., andFarstad, M. 1979. Intracellular Localization of Long-Chain Acyl-CoA Hydrolase and Acyl-Carnitine Hydrolase in Brown Adipose Tissue from Guinea Pigs. Biochem. J. 182:347–352.

    Google Scholar 

  6. Knauer, T. E., Gureck, J. J., andKnauer, G. R. 1980. Substrate Stabilization of the Palmitoyl-Coenzyme A Hydrolase Activity of Rat Submaxillary Gland. Biochem. J. 187:269–272.

    Google Scholar 

  7. Berge, R. K. 1979. Purification and Characterization of a Long-Chain Acyl-CoA Hydrolase from Rat Liver Microsomes. Biochim. Biophys. Acta 574:321–333.

    Google Scholar 

  8. Srere, P. A., Seubert, W., andLynen, F. 1959. Palmityl Coenzyme A Deacylase. Biochim. Biophys. Acta 33:313–319.

    Google Scholar 

  9. Anderson, A. D., andErwin, V. G. 1971. Brain Acyl-Coenzyme A Hydrolase: Distribution, Purification and Properties. J. Neurochem. 18:1179–1186.

    Google Scholar 

  10. Knauer, T. E. 1979. Factors Affecting the Activity and Stability of the Palmitoyl-Coenzyme A Hydrolase of Rat Brain. Biochem. J. 179:515–523.

    Google Scholar 

  11. Porter, J. W., andLong, R. W. 1958. A Study of the Role of Palmityl Coenzyme A in Fatty Acid Synthesis by the Pigeon Liver System. J. Biol. Chem. 233:20–31.

    Google Scholar 

  12. McMurray, W. C., andMagee, W. L. 1972. Phospholipid Metabolism. Ann. Rev. Biochem. 41:129–160.

    Google Scholar 

  13. Wieland, O., Weiss, L., andEger-Neufeldt, I. 1964. Hemmung der Enzymatischen Citronensauersynthese durch Langkettige Acyl-Thioester des Coenzym A. Biochem. Z. 339:501–513.

    Google Scholar 

  14. Srere, P. A. 1965. Palmitoyl-Coenzyme A Inhibition of the Citate Condensing Enzyme. Biochim. Biophys. Acta 106:445–455.

    Google Scholar 

  15. Taketa, K., andPogell, B. M. 1966. The Effect of Palmityl Coenzyme A on Glucose 6-Phosphate Dehydrogenase and Other Enzymes. J. Biol. Chem. 241:720–726.

    Google Scholar 

  16. Goodridge, A. G. 1973. Regulation of Fatty Acid Synthesis in Isolated Hepatocytes. Evidence for a Physiological Role for Long Chain Acyl Coenzyme A and Citrate. J. Biol. Chem. 248:4318–4326.

    Google Scholar 

  17. Kawaguchi, A., andBloch, K. 1974. Inhibition of Glucose-6-Phosphate Dehydrogenase by Palmitoyl Coenzyme A. J. Biol. Chem. 249:5793–5800.

    Google Scholar 

  18. Hsu, K. L., andPowell, G. L. 1975. Inhibition of Citrate Synthease by Oleoyl-CoA: A Regulatory Phenomenon. Proc. Nat. Acad. Sci. USA 72:4729–4733.

    Google Scholar 

  19. Kawaguchi, A., andBloch, K. 1976. Inhibition of Glutamate Dehydrogenase and Malate Dehydrogenase by Palmitoyl Coenzyme A. J. Biol. Chem. 251:1406–1412.

    Google Scholar 

  20. Nikawa, J., Tanabe, T., Ogiwara, H., Shiba, T., andNuma, S. 1979. Inhibitory Effects of Long-Chain Acyl Coenzyme A Analogues on Rat Liver Acetyl Coenzyme A Carboxylase. FEBS Lett 102:223–228.

    Google Scholar 

  21. Mita, M., andYasumasu, I. 1981. Inhibition of Dihydrofolate Reductase by Palmitoyl Coenzyme A. Int. J. Biochem. 13:229–232.

    Google Scholar 

  22. Vignais, P. V. 1976. Molecular and Physiological Aspects of Adenine Nucleotide Transport in Mitochondria. Biochim. Biophys. Acta 456:1–38.

    Google Scholar 

  23. Asimakis, G. K., andSordahl, L. A. 1977. Effects of Atractyloside and Palmitoyl Coenzyme A on Calcium Transport in Cardiac Mitochondria. Arch. Biochem. Biophys. 179:200–210.

    Google Scholar 

  24. Wood, J. M., Bush, B., Pitts, B. J. R., andSchwartz, A. 1977. Inhibition of Bovine Heart Na+, K+-ATPase by Palmitylcarnitine and Palmityl-CoA. Biochem. Biophys. Res. Commun. 74:677–684.

    Google Scholar 

  25. Lee, K. Y., andSchulz, H. 1979. Isolation, Properties, and Regulation of a Mitochondrial Acyl Coenzyme A Thioesterase from Pig Heart. J. Biol. Chem. 254:4516–4523.

    Google Scholar 

  26. Sun, G. Y., Smith, R. E., Chan, K., andMacQuarrie, R. 1980. Inhibition of Acyl-CoA Hydrolase Activity in Liver Microsomes by Lyso-Phospholipids. Biochem. Biophys. Res. Commun. 94:1278–1284.

    Google Scholar 

  27. Ellman, G. L. 1959. Tissue Sulfhydryl Groups. Arch. Biochem. Biophys. 82:70–77.

    Google Scholar 

  28. Glazer, A. N., DeLange, R. J., andSigman, D. S. 1975. Pages 113,in Work, T. S., andWork, E., (eds.), Chemical Modification of Proteins.

  29. Bonnor, W. M., andBlock, K. 1972, Purification and Properties of Fatty Acyl Thioesterase I from Escherichia Coli. J. Biol. Chem. 247:3123–3133.

    Google Scholar 

  30. Lee, K. Y., andSchulz, H. 1979. Isolation Properties, and Regulation of a Mitochondrial Acyl Coenzyme A Thioesterase from Pig Heart. J. Biol. Chem. 254:4516–4523.

    Google Scholar 

  31. Libertine, L. J., andSmith, S. 1978., Purification and Properties of a Thioesterase from Lactating Rat Mammary Gland which Modifies the Product Specificity of Fatty Acid Synthetase. J. Biol. Chem. 253:1393–1401.

    Google Scholar 

  32. Kezdy, F. J., andBender, M. L. 1962. The Kinetics of the Chymotrypsin-Catalyzed Hydrolysis of p-Nitrophenyl Acetate. Biochem. 6:1097–1106.

    Google Scholar 

  33. Bradford, M. 1976. A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Anal. Biochem. 72:248254.

    Google Scholar 

  34. Ma, P. F., Betras, S., andDunnington, G. 1976. Rapid and Efficient Separation and Identification of the Two Molecular Forms of Human Adenosine Deaminase by Thin-Layer Gel Filtration Chromatography. Anal. Biochem. 75:177–182.

    Google Scholar 

  35. MacQuarrie, R., andBuel 1982. Physical and Catalytic Properties of the Isozymes of Adenosine Deaminase from Human Red Blood Cells. Molec. Cell Biochem. 48:121–126.

    Google Scholar 

  36. Tiselius, A., Hjerten, S., andLevin, O. 1956. Protein Chromatography on Calcium Phosphate Columns. Arch. Biochem. Biophys. 65:132–155.

    Google Scholar 

  37. Mazin, A. L., Sulimova, G. E., andVanyuskin, B. F. 1974. Granulated Hydroxyapatite: Preparation and Chromatographic Properties. Anal. Biochem. 61:62–71.

    Google Scholar 

  38. Davis, B. J. 1965. Disc Electrophoresis—II Method and Application to Human Serum Proteins. Ann. N. Y. Acad. Sci. 121:404–427.

    Google Scholar 

  39. Weber, K., andOsborn, M. 1969. The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis. J. Biol. Chem. 244:4406–4412.

    Google Scholar 

  40. Fairbank, G., Steck, T. L., andWallach, D. F. H. 1971. Electrophoretic Analysis of the Major Polypeptides of the Human Erythrocyte Membrane. Biochem. 10:2606–2617.

    Google Scholar 

  41. Berge, R. K., andFarstad, M. 1976. Purification and Characterization of Long-Chain Acyl-CoA Hydrolase from Rat Liver Mitochondria. Eur. J. Biochem. 96:393–401.

    Google Scholar 

  42. Gross, R. W. 1983. Purification of Rabbit Myocardial Cytosolic Acyl-CoA Hydrolase, Identity with Lysophospholipase, and Modulation of Enzymic Activity by Endogenous Cardiac Amphiphiles. Biochem. 22:5641–5646.

    Google Scholar 

  43. Berge, R. K. 1980. Physiochemical Properties of the Long-Chain-Acyl-CoA Hydrolase from Rat Liver Microsomes. Eur. J. Biochem. 111:67–72.

    Google Scholar 

  44. Bonser, R. W., andLunt, G. G. 1976. The Incorporation [1-14C] Acetate into Unesterified Fatty Acids in Rat Cerebral Cortex in vivo. J. Neurochem. 26:331–334.

    Google Scholar 

  45. Buttlaire, D. H., Cohn, M., andBridger, W. A. 1977. Interactions of Phospho- and Dephosphosuccinyl Coenzyme A Synthetase with Manganous Ion and Substrates. J. Biol. Chem. 252:1957–1964.

    Google Scholar 

  46. Fischer, B. F., Haring, U. K., Tribolet, R., andSigel, H. 1979. Metal Ion/Buffer Interactions Stability of Binary and Ternary Complexes containing 2-Amino-2(Hydroxymethyl)-1,3-Propanediol (Tris) and Adenosine-5′-Triphosphate (ATP). Eur. J. Biochem. 94:523–530.

    Google Scholar 

  47. Berge, R. K., andDossland, B. 1979. Differences between Microsomal and Mitochondrial-Matrix Palmitoyl-Coenzyme A Hydrolase, and Palmitoyl-L-Carnitine Hydrolase from Rat Liver. Biochem. J. 181:119–125.

    Google Scholar 

Download references

Author information

Author notes

  1. Augustine Y. Lin

    Present address: Department of Chemistry, University of Southern California, 90089, Los Angeles, California

Authors and Affiliations

  1. Department of Chemistry and School of Medicine, University of Missouri-Kansas City, 64110, Kansas City, Missouri

    Ron macQuarrie

  2. Sinclair Research Farm and Biochemistry Department, University of Missouri, 65201, Columbia, Missouri

    Grace Y. Sun

Authors
  1. Augustine Y. Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Grace Y. Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ron macQuarrie
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lin, A.Y., Sun, G.Y. & macQuarrie, R. Partial purification and properties of long-chain acyl-CoA hydrolase from rat brain cytosol. Neurochem Res 9, 1571–1591 (1984). https://doi.org/10.1007/BF00964592

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00964592

Keywords

Search

Navigation