Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Jul 15;285(Pt 2):401–404. doi: 10.1042/bj2850401

Relationship between glutathione content in liver and glutathione conjugation rate in the rat in vivo. Effect of buthionine sulphoximine pretreatment on conjugation of the two 2-bromoisovalerylurea enantiomers during intravenous infusion.

M Polhuijs 1, G Lankhaar 1, G J Mulder 1
PMCID: PMC1132802  PMID: 1637334

Abstract

The relationship between hepatic glutathione content and hepatic glutathione conjugation rate in the rat in vivo was investigated. As substrate for glutathione conjugation, racemic (R,S)-2-bromoisovalerylurea (BIU) was used which gives rise to the biliary excretion of two diastereoisomeric glutathione conjugates and the urinary excretion of two diastereoisomeric mercapturates. The excretion rate of the glutathione conjugate in bile reflects hepatic conjugation exclusively. An intravenous infusion of BIU was given and the excretion rates of the metabolites in bile and urine were determined. The glutathione concentration in the liver was followed by taking biopsies every hour. Glutathione was depleted by the infused substrate; in rats that were pretreated with the inhibitor of glutathione biosynthesis, buthionine sulphoximine (BSO), the depletion of the glutathione content was more rapid. The rate of excretion of the glutathione conjugate in bile was plotted against hepatic glutathione content. These results indicate that the 'organ Km' for glutathione in the liver is approximately 0.5 mumol/g of liver, so that the hepatic glutathione conjugation rate is decreased only at severe glutathione depletion.

Full text

PDF
401

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adang A. E., Brussee J., Meyer D. J., Coles B., Ketterer B., van der Gen A., Mulder G. J. Substrate specificity of rat liver glutathione S-transferase isoenzymes for a series of glutathione analogues, modified at the gamma-glutamyl moiety. Biochem J. 1988 Oct 15;255(2):721–724. [PMC free article] [PubMed] [Google Scholar]
  2. Akerboom T. P., Sies H. Assay of glutathione, glutathione disulfide, and glutathione mixed disulfides in biological samples. Methods Enzymol. 1981;77:373–382. doi: 10.1016/s0076-6879(81)77050-2. [DOI] [PubMed] [Google Scholar]
  3. Anderson M. E. Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol. 1985;113:548–555. doi: 10.1016/s0076-6879(85)13073-9. [DOI] [PubMed] [Google Scholar]
  4. Chen W. J., Graminski G. F., Armstrong R. N. Dissection of the catalytic mechanism of isozyme 4-4 of glutathione S-transferase with alternative substrates. Biochemistry. 1988 Jan 26;27(2):647–654. doi: 10.1021/bi00402a023. [DOI] [PubMed] [Google Scholar]
  5. D'Silva C. Inhibition and recognition studies on the glutathione-binding site of equine liver glutathione S-transferase. Biochem J. 1990 Oct 1;271(1):161–165. doi: 10.1042/bj2710161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Griffith O. W. Depletion of glutathione by inhibition of biosynthesis. Methods Enzymol. 1981;77:59–63. doi: 10.1016/s0076-6879(81)77011-3. [DOI] [PubMed] [Google Scholar]
  7. Ivanetich K. M., Goold R. D. A rapid equilibrium random sequential bi-bi mechanism for human placental glutathione S-transferase. Biochim Biophys Acta. 1989 Sep 14;998(1):7–13. doi: 10.1016/0167-4838(89)90111-8. [DOI] [PubMed] [Google Scholar]
  8. Mannervik B., Danielson U. H. Glutathione transferases--structure and catalytic activity. CRC Crit Rev Biochem. 1988;23(3):283–337. doi: 10.3109/10409238809088226. [DOI] [PubMed] [Google Scholar]
  9. Mannervik B. The isoenzymes of glutathione transferase. Adv Enzymol Relat Areas Mol Biol. 1985;57:357–417. doi: 10.1002/9780470123034.ch5. [DOI] [PubMed] [Google Scholar]
  10. Mulder G. J., Scholtens E., Meijer D. K. Collection of metabolites in bile and urine from the rat. Methods Enzymol. 1981;77:21–30. doi: 10.1016/s0076-6879(81)77006-x. [DOI] [PubMed] [Google Scholar]
  11. Plummer J. L., Smith B. R., Sies H., Bend J. R. Chemical depletion of glutathione in vivo. Methods Enzymol. 1981;77:50–59. doi: 10.1016/s0076-6879(81)77010-1. [DOI] [PubMed] [Google Scholar]
  12. Polhuijs M., Kuipers F., Vonk R. J., Mulder G. J. Stereoselectivity of glutathione conjugation: blood elimination of alpha-bromoisovalerylurea enantiomers and biliary excretion of the conjugates in unanesthetized normal or congenitally jaundiced rats. J Pharmacol Exp Ther. 1989 Jun;249(3):874–878. [PubMed] [Google Scholar]
  13. Polhuijs M., Meijer D. K., Mulder G. J. The fate of diastereomeric glutathione conjugates of alpha-bromoisovalerylurea in blood in the rat in vivo and in the perfused liver. Stereoselectivity in biliary and urinary excretion. J Pharmacol Exp Ther. 1991 Feb;256(2):458–461. [PubMed] [Google Scholar]
  14. Sedlak J., Lindsay R. H. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 1968 Oct 24;25(1):192–205. doi: 10.1016/0003-2697(68)90092-4. [DOI] [PubMed] [Google Scholar]
  15. Te Koppele J. M., Coles B., Ketterer B., Mulder G. J. Stereoselectivity of rat liver glutathione transferase isoenzymes for alpha-bromoisovaleric acid and alpha-bromoisovalerylurea enantiomers. Biochem J. 1988 May 15;252(1):137–142. doi: 10.1042/bj2520137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. te Koppele J. M., Dogterom P., Vermeulen N. P., Meijer D. K., van der Gen A., Mulder G. J. alpha-Bromoisovalerylurea as model substrate for studies on pharmacokinetics of glutathione conjugation in the rat. II. Pharmacokinetics and stereoselectivity of metabolism and excretion in vivo and in the perfused liver. J Pharmacol Exp Ther. 1986 Dec;239(3):905–914. [PubMed] [Google Scholar]
  17. te Koppele J. M., de Lannoy I. A., Pang K. S., Mulder G. J. Stereoselective glutathione conjugation and amidase-catalyzed hydrolysis of alpha-bromoisovalerylurea enantiomers in isolated rat hepatocytes. J Pharmacol Exp Ther. 1987 Oct;243(1):349–355. [PubMed] [Google Scholar]
  18. te Koppele J. M., van der Mark E. J., Mulder G. J. Liquid chromatographic determination of diastereomeric glutathione conjugates and further derivatives of alpha-bromoisovalerylurea in rat bile and urine by electrochemically generated bromine. J Chromatogr. 1988 May 13;427(1):67–77. doi: 10.1016/0378-4347(88)80105-1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES