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. 1988 Jan 1;249(1):239–245. doi: 10.1042/bj2490239

The soluble carnitine palmitoyltransferase from bovine liver. A comparison with the enzymes from peroxisomes and from the mitochondrial inner membrane.

R R Ramsay 1
PMCID: PMC1148690  PMID: 3124822

Abstract

The properties of two carnitine acyltransferases (CPT) purified from bovine liver are compared to confirm that they are different proteins. The soluble CPT and the inner CPT from mitochondria differ in subunit Mr, native Mr, pI and reactivity with thiol reagents. All eight free thiol groups in soluble CPT react with 5,5'-dithiobis-(2-nitrobenzoate) in the absence of any unfolding reagent, and activity is gradually lost. The inner CPT activity is completely stable in the presence of 5,5'-dithiobis-(2-nitrobenzoate), and only one thiol group per molecule of subunit is modified in the native enzyme. Antisera to each enzyme inhibit that enzyme, but do not cross-react. CPT activity in subcellular fractions can now be identified by titration with these antibodies. The soluble CPT from bovine liver is probably peroxisomal in origin, but, although antigenically similar, it differs from the peroxisomal carnitine octanoyltransferase found in rat and mouse liver in its specificity for the longer-chain acyl-CoA substrates.

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Selected References

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  1. Bergstrom J. D., Reitz R. C. Studies on carnitine palmitoyl transferase: the similar nature of CPTi (inner form) and CPTo (outer form). Arch Biochem Biophys. 1980 Oct 1;204(1):71–79. doi: 10.1016/0003-9861(80)90008-9. [DOI] [PubMed] [Google Scholar]
  2. Brady P. S., Dunker A. K., Brady L. J. Characterization of hepatic carnitine palmitoyltransferase. Use of bromoacyl derivatives and antibodies. Biochem J. 1987 Feb 1;241(3):751–757. doi: 10.1042/bj2410751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  4. Clarke P. R., Bieber L. L. Isolation and purification of mitochondrial carnitine octanoyltransferase activities from beef heart. J Biol Chem. 1981 Oct 10;256(19):9861–9868. [PubMed] [Google Scholar]
  5. Declercq P. E., Falck J. R., Kuwajima M., Tyminski H., Foster D. W., McGarry J. D. Characterization of the mitochondrial carnitine palmitoyltransferase enzyme system. I. Use of inhibitors. J Biol Chem. 1987 Jul 15;262(20):9812–9821. [PubMed] [Google Scholar]
  6. Farrell S. O., Fiol C. J., Reddy J. K., Bieber L. L. Properties of purified carnitine acyltransferases of mouse liver peroxisomes. J Biol Chem. 1984 Nov 10;259(21):13089–13095. [PubMed] [Google Scholar]
  7. Ghosh M. K., Hajra A. K. A rapid method for the isolation of peroxisomes from rat liver. Anal Biochem. 1986 Nov 15;159(1):169–174. doi: 10.1016/0003-2697(86)90323-4. [DOI] [PubMed] [Google Scholar]
  8. Healy M. J., Kerner J., Bieber L. L. Enzymes of carnitine acylation. Is overt carnitine palmitoyltransferase of liver peroxisomal carnitine octanoyltransferase? Biochem J. 1988 Jan 1;249(1):231–237. doi: 10.1042/bj2490231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoppel C. L., Tomec R. J. Carnitine palmityltransferase. Location of two enzymatic activities in rat liver mitochondria. J Biol Chem. 1972 Feb 10;247(3):832–841. [PubMed] [Google Scholar]
  10. Ishii S., Ishii H., Suga T. The presence of peroxisomal carnitine palmitoyltransferase in chick embryo liver. J Biochem. 1985 Sep;98(3):747–755. doi: 10.1093/oxfordjournals.jbchem.a135332. [DOI] [PubMed] [Google Scholar]
  11. Leighton F., Poole B., Beaufay H., Baudhuin P., Coffey J. W., Fowler S., De Duve C. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968 May;37(2):482–513. doi: 10.1083/jcb.37.2.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Markwell M. A., McGroarty E. J., Bieber L. L., Tolbert N. E. The subcellular distribution of carnitine acyltransferases in mammalian liver and kidney. A new peroxisomal enzyme. J Biol Chem. 1973 May 25;248(10):3426–3432. [PubMed] [Google Scholar]
  13. McGarry J. D., Leatherman G. F., Foster D. W. Carnitine palmitoyltransferase I. The site of inhibition of hepatic fatty acid oxidation by malonyl-CoA. J Biol Chem. 1978 Jun 25;253(12):4128–4136. [PubMed] [Google Scholar]
  14. Miyazawa S., Ozasa H., Osumi T., Hashimoto T. Purification and properties of carnitine octanoyltransferase and carnitine palmitoyltransferase from rat liver. J Biochem. 1983 Aug;94(2):529–542. doi: 10.1093/oxfordjournals.jbchem.a134384. [DOI] [PubMed] [Google Scholar]
  15. Murthy M. S., Pande S. V. Malonyl-CoA binding site and the overt carnitine palmitoyltransferase activity reside on the opposite sides of the outer mitochondrial membrane. Proc Natl Acad Sci U S A. 1987 Jan;84(2):378–382. doi: 10.1073/pnas.84.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ramsay R. R., Derrick J. P., Friend A. S., Tubbs P. K. Purification and properties of the soluble carnitine palmitoyltransferase from bovine liver mitochondria. Biochem J. 1987 Jun 1;244(2):271–278. doi: 10.1042/bj2440271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. TABOR C. W., TABOR H., ROSENTHAL S. M. Purification of amine oxidase from beef plasma. J Biol Chem. 1954 Jun;208(2):645–661. [PubMed] [Google Scholar]
  18. West D. W., Chase J. F., Tubbs P. K. The separation and properties of two forms of carnitine palmitoyltransferase from ox liver mitochondria. Biochem Biophys Res Commun. 1971 Mar 5;42(5):912–918. doi: 10.1016/0006-291x(71)90517-1. [DOI] [PubMed] [Google Scholar]
  19. Woeltje K. F., Kuwajima M., Foster D. W., McGarry J. D. Characterization of the mitochondrial carnitine palmitoyltransferase enzyme system. II. Use of detergents and antibodies. J Biol Chem. 1987 Jul 15;262(20):9822–9827. [PubMed] [Google Scholar]
  20. Zammit V. A., Corstorphine C. G. Altered release of carnitine palmitoyltransferase activity by digitonin from liver mitochondria of rats in different physiological states. Biochem J. 1985 Sep 1;230(2):389–394. doi: 10.1042/bj2300389. [DOI] [PMC free article] [PubMed] [Google Scholar]

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