Skip to main content
SpringerLink
Log in

Purification of carnitine acetyltransferase from skeletal muscle of the camel (Camelus dromedarius)

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The enzyme carnitine acetyltransferase (CAT) catalyzes the reversible transfer of short-chain acyl groups between coenzyme A and L-carnitine, and hence, plays an important role in the β-oxidation of fatty acids. Purification and characterization of CAT from desert animal species may help in explaining the involvement of secondary pathways for energy production in these species. In this paper, we report the purification and partial characterization of CAT from the Arabian camel. CAT was purified from the skeletal muscle of the Arabian camel by ammonium sulfate and acetone fractionation, followed by chromatography on DEAF-Sepharose, agarose-Co A and Superose 12 gel filtration columns. CAT was purified by 2937-fold to a specific activity of 94 Units mg−1. The purified CAT was a monomer of 59 kDa as judged by native and SDS-PAGE, and showed a pl of 5.2. The enzyme displayed maximum activity with propionyl-Co A. Apparent Km for acetyl-, propionyl- and butyryl-Co A were 27.7, 17.3 and 29 μM respectively, while palmitoyl-Co A was not a substrate.

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. Bremer J: Carnitine-metabolism and functions. Physiol Rev 63: 1420–1480, 1983

    Google Scholar 

  2. Bieber LL: Carnitine. Ann Rev Biochem. 57: 261–283, 1988

    Google Scholar 

  3. Bieber LL, Lysiak W: Characteristics and functions of short-chain and medium-chain carnitine acyltransferases. In: P. R. Borum (eds). Clinical aspects of human carnitine deficiency. Pergamon Press, New York, 1986, pp 157–164

    Google Scholar 

  4. Hoppel C: The physiological role of L-carnitine. In: R Ferrari, S. DiMauro and G. Sherwood (eds). L-carnitine and its role in medicine: From functions to therapy. Academic Press, California, 1992, pp 5–19

    Google Scholar 

  5. Chase JFA, Pearson DJ, Tubbs PK: The purification of crystalline carnitine acetyltransferase. Biochem Biophys Acta 96: 162–165, 1965

    Google Scholar 

  6. Farrel SO, Fiol CJ, Reddy JK, Bieber LL: Properties of purified carnitine acetyltransferases of mouse liver peroxisomes. J Biol Chem 259: 13089–13095, 1984

    Google Scholar 

  7. Chung C, Chung CD, Bieber LL: Purification of heart and liver mitochondrial carnitine acetyltransferase. Protein Exp Pur 2: 426–431, 1991

    Google Scholar 

  8. Bliosi W, Colombo I, Garavaglia B, Giardini R, Finocchiaro G, Didonato S: Purification and properties of carnitine acetyltransferase from human liver. Eur J Biochem 189: 529–546, 1990

    Google Scholar 

  9. Fritz IB, Schultz SK, Srere PA: Properties of partially purified carnitine acetyltransferase. J Biol Chem 238: 2509–2517, 1963

    Google Scholar 

  10. Chase JFA: The substrate specificity of carnitine acetyltransferase. Biochem. J 104: 510–518, 1967

    Google Scholar 

  11. Miyazawa S, Ozasa H, Furuta S, Osumi T, Hashimoto T: Purification and properties of carnitine acetyltransferase from rat liver. J Biochem 93: 439–451, 1983

    Google Scholar 

  12. MacFarlane WV, Morris RJH, Howard B: Turn-over and distribution of water in desert camels, sheep, cattle and kangaroos. Nature 197: 270–271, 1963

    Google Scholar 

  13. Yagil R Sod-Moriah UA, Meyerstein N: Dehydration and camel blood. I. The life span of the camel erythrocyte. Am J Physiol 226: 298–301, 1974

    Google Scholar 

  14. Hussein MF, Al-Momen AK, Gader AMA: Haemostatic parameters in the camel (Camelus dromedarius): comparison with humans. Comp Haematol Int 2: 92–96, 1992

    Google Scholar 

  15. Yagil R: In Comparative Animal Nutrition Vol 5, The Desert Camel: Comparative Physiological Adaptation. S. Karger, Switzerland, 1985, pp 10–42

    Google Scholar 

  16. Snow DH, Billah A, Ridha A: Effects of maximal exercise on the blood composition of the racing camel. Vet Rec 123: 311–312, 1988

    Google Scholar 

  17. Rose RJ, Evans DL, Henckel P, Knight PK, Cluer D, Saltin B: Metabolic responses to prolonged exercise in the racing camel. Acta Physiol Scand Suppl 617: 49–60, 1994

    Google Scholar 

  18. Alhomida AS, Duhaiman AS, Al-Jafari AA, Junaid MA: Determination of L carnitine, acylcarnitine and total carnitine levels in plasma and tissues of camel (Camelus dromedarius). Comp Biochem Physiol 111 B: 441–445, 1995

    Google Scholar 

  19. Chase JFA: Carnitine acetyltransferase from pigeon breast muscle. Methods Enzmol 13: 387–393, 1969

    Google Scholar 

  20. Mittal B, Kurup CKR: Purification of clofibrate-induced carnitine acetyltransferase from rat liver mitochondria. Biochem Biophys Acta 619: 90–97, 1980

    Google Scholar 

  21. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275, 1951

    CAS  PubMed  Google Scholar 

  22. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  23. Duhaiman AS, Rabbani N, Cotlier E: Camel lens crystallin glycosylation and high molecular weight aggregate formation in the presence of ferrous ions and glucose. Biochem Biophys Res Commun 173: 823–832, 1990

    Google Scholar 

  24. Saltin B, Rose TtJ, Henckel P: Skeletal muscle morphology and metabolic potential. Acta Physiol Scand suppl 617: 24–32, 1994

    Google Scholar 

  25. Rose RJ, Purdue RA, Hensley W: Plasma biochemistry alterations in horses during an endurance ride. Equine vet J 9: 122–126, 1977

    Google Scholar 

  26. Shephard RJ, Kavanagh TJ: Biochemical changes with marathon running. Observation on post coronary patients. In: H. Howald and J.R Poortmans (eds). Metabolic Adaptations to prolonged physical exercise. Birkhauser verlag, Basel, Switzerland, 1975, pp 245–252

    Google Scholar 

  27. Snow DH, Kerr MG, Nimmo MA, Abbott EM: Alterations in blood, sweat, urine and muscle composition during prolonged exercise in the horse. Vet Rec 10, 377–384, 1982

    Google Scholar 

  28. Clarke PRK, Bieber LL: Isolation and purification of mitochondrial carnitine octanoyltransferase activities from beefheart. J Biol Chem 256: 9861–9868, 1981

    Google Scholar 

  29. Markwell MK, Tolbert NE, Bieber LL: Comparision of the acetyltransferase activities from rat liver peroxisomes and microsomes. Arch Biochem Biophys 176: 479–188, 1976

    Google Scholar 

  30. Chase JFA, Tubbs PK: Some kinetic studies on the mechanism of action of carnitine acetyltransferase. Biochem J 99: 32–40, 1966

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, College of Science, King Saud University, P.O. Box 2455, 11451, Riyadh, Saudi Arabia

    A. S. Alhomida, A. S. Duhaiman, A. A. Al-Jafari & M. A. Junaid

Authors
  1. A. S. Alhomida
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. S. Duhaiman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. A. Al-Jafari
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. A. Junaid
    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

Alhomida, A.S., Duhaiman, A.S., Al-Jafari, A.A. et al. Purification of carnitine acetyltransferase from skeletal muscle of the camel (Camelus dromedarius). Mol Cell Biochem 165, 95–101 (1996). https://doi.org/10.1007/BF00229470

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00229470

Key words

Search

Navigation