Comparison of different activators and diazotization procedures in the assay of bilirubin-UDP-glucuronyl transferase activity in rat liver

Clinica Chimica Acta; International Journal of Clinical Chemistry
N TavoloniP D Berk

Abstract

The activity of bilirubin-UDP-glucuronyl transferase (UDPGT) in rat liver microsomes was assayed in standardized incubation mixtures employing two different procedures to quantitate formation of conjugated bilirubin: solvent extraction and diazotization with sulfanilic acid, or selective coupling with diazotized ethyl anthranilate at pH 2.7. Both of these procedures yielded equivalent UDPGT activities with several different microsomal preparations, averaging, respectively, 385 and 372 microU protein with untreated microsomes, 646 and 608 microU protein following activation with UDP-N-acetylglucosamine (3.07 mmol/l), and 1370 and 1478 microU protein after treatment with digitonin (0.65%). In rats pretreated with phenobarbital (75 mg/kg per 6 days), a 70-90% increase in UDPGT activity was observed with either diazotization procedure, irrespective of whether 'native', UDP-N-acetylglucosamine-or digitonin-activated enzyme was employed. These studies indicate that, when other conditions are standardized, equivalent information about UDPGT can be obtained with either of two widely employed diazotization reactions and a variety of enzyme activation procedures.

References

Nov 10, 1972·Biochimica Et Biophysica Acta·A Winsnes
Nov 4, 1969·Biochimica Et Biophysica Acta·A Winsnes
Nov 1, 1970·The Biochemical Journal·J D Ostrow, N H Murphy
Jul 1, 1970·Clinica Chimica Acta; International Journal of Clinical Chemistry·M BlackK P Heirwegh
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Nov 1, 1971·FEBS Letters·A F. McDonagh, F Assisi
Nov 1, 1962·Clinica Chimica Acta; International Journal of Clinical Chemistry·A P WEBER, L SCHALM

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Citations

Jan 1, 1991·The International Journal of Biochemistry·R A FreedlandC E Cornelius
Jan 1, 1983·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·K D Cole, G H Little
Jan 1, 1990·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·L S Fang, M S Hung

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