Abstract
In experiments with 4 different types of cells, we evaluated the cholesterogenic activity by incorporation of14C-acetic acid into cholesterol and digitonin-precipitable sterols. In every case, the cholesterogenesis appeared considerably faster when expressed as digitonid than when expressed as real cholesterol production, and sometimes the data obtained by the 2 methods were contradictory. Detailed analysis of both digitonid components and nonprecipitable radioactive metabolites showed that a very variable fraction of methyl sterols (including bifunctional methyl sterols) co-precipitates with the C-27 sterols. In cholesterol regulation studies and particularly when the cells exhibit a low cholesterogenesis, the digitonin method is unsuitable and can lead to erroneous interpretations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- TLC:
-
thin layer chromatography
- GLC:
-
gas liquid chromatography
- 4,4′-dimethyl sterols or C-29 sterols:
-
cholesten-4,4′ dimethyl-3β-ols
- 4α-methyl sterols or C-28 sterols:
-
cholesten-4α-methyl-3β-ols
- HMG CoA:
-
3-hydroxy-3-methyl-glutaryl coenzyme A
References
Kandutsch, A.A., and Sauceer, E. (1969) J. Biol. Chem. 244, 2299–2306.
Philippot, J.R., Cooper, A.G., and Wallach, D.F.H. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 956–960.
Philippot, J.R., Cooper, A.G., and Wallach, D.F.H. (1975) Biochim. Biophys. Acta 406, 161–166.
Jeske, D.J., and Dietschy, J.M. (1980) J. Lipid Res. 20, 740–752.
Picard, F., Duval, D., and Homo, F. (1981) J. Steroid Biochem. 15, 461–466.
Heiniger, H.J., and Marshall, J.D. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 3823–3827.
Cenedella, R.J. (1982) Lipids 17, 443–447.
Tabacik, C., Astruc, M., Laporte, M., Descomps, B., and Crastes de Paulet, A. (1979) Biochem. Biophys. Res. Commun. 88, 706–712.
Tabacik, C., Astruc, M., Laporte, M., Descomps, B., Crastes de Paulet, A., and Serrou, B. (1981) Biomedecine 34, 128–132.
Sultan, C., Migeon, B.R., Rothwell, S.W., Maes, M., and Migeon, C.J. (1980) Pediatr. Res. 13, 67–69.
Sperry, W.M., and Webb, M. (1950) J. Biol. Chem. 187, 97–106.
Tabacik, C., Aliau, S., Serrou, B., and Crastes de Paulet, A. (1981) Biochem. Biophys. Res. Commun. 101, 1087–1095.
Bradford, M. (1976) Anal. Biochem. 72, 248.
Tabacik, C., and Aliau, S. 24th International Conference on the Biochemistry of Lipids, Toulouse, September 14–16, 1983.
Tabacik, C., Aliau, S., Devilliers, C., and Sultan, C., 30th meeting of the European Tissue Culture Society, Paris, France. July 15–17, 1982.
Author information
Authors and Affiliations
About this article
Cite this article
Tabacik, C., Aliau, S. & Crastes de Paulet, A. Digitonin-precipitable sterols as a measure of cholesterol biosynthesis: Contradictory results. Lipids 18, 641–649 (1983). https://doi.org/10.1007/BF02534676
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02534676