Quantitative studies of sulphate conjugation by isolated rat liver cells using [35S]sulphate.
We have developed a simple, rapid and sensitive method for the study of sulphate conjugation in isolated liver cells based on the incorporation of 35S from [35S]sulphate. Excess [35S]sulphate is removed by a barium precipitation procedure, leaving [35S]sulphate conjugates in solution. We have used this method to examine the kinetics of sulphation of N-acetyl-p-aminophenol (acetaminophen), 4-nitrophenol and 1-naphthol in isolated rat liver cells. The efficiency of recovery of the sulphate conjugates was greater than 86%. The method is applicable to the quantitative study of sulphate conjugation of any substrate which forms a sulphate conjugate that is soluble in the presence of barium, without the need for standards or radiolabelled sulphate acceptors.
Dawson
J
J
Biochemical Sciences, Wellcome Research Laboratories, Beckenham, Kent, U.K.
Knowles
R G
RG
Pogson
C I
CI
D000082:Acetaminophen / Q000378:metabolism; D000818:Animals; D001464:Barium; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D005965:Glucuronates / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D009284:Naphthols / Q000378:metabolism; D009596:Nitrophenols / Q000378:metabolism; D051381:Rats; D013431:Sulfates / Q000378:metabolism*; D013462:Sulfur Radioisotopes / Q000032:analysis
D005965:Glucuronates; D009284:Naphthols; D009596:Nitrophenols; D013431:Sulfates; D013462:Sulfur Radioisotopes; D001464:Barium; C029350:1-naphthol; D000082:Acetaminophen; C024836:4-nitrophenol