An improved assay for steroid glucuronyltransferase in rat liver microsomes

Rao, Govind S.; Haueter, Gudrun; Rao, Marie Luise; Breuer, H.

doi:10.1016/0003-2697(76)90307-9

Cited by 78 publications

(24 citation statements)

References 7 publications

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“…Microsomes which had been frozen and thawed once only were used. Established assay procedures for the following aglycones were used: 2-aminophenol [15]; 1-naphthol [16]; testoster one and androsterone [17]; bilirubin [18], and 5-hydroxytryptamine [19]. Protein in microsomal sam ples was determined by the method of Lowry et al…”

Section: Methodsmentioning

confidence: 99%

Development of Human Liver UDP-Glucuronosyltransferases

Burchell¹,

Coughtrie²,

Jackson³

et al. 1989

Dev Pharmacol Ther

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The development of multiple UDPGT activities towards eight substrates has been studied in fetal term and adult post-mortem (less than 5 h after death) liver samples. Most fetal and term liver activities were less than 14% of adult values, except that towards 5-hydroxytryptamine which was present in fetal and term liver at adult levels. The majority of UDPGT activities develop to adult levels within 10-20 weeks postnatally, and even premature (30 weeks) which survive for up to 10 weeks will develop these enzyme activities. Immunoblot analysis of human liver microsomes and cDNA cloning of human UDPGT shows the existence of the family of isoenzymes in man, and it is important to determine the developmental pattern of individual drug glucuronidating enzymes in liver. Immunoblot analysis of developing liver shows the presence of two major UDPGT polypeptides in fetal liver, whereas more than five are observed in adult liver. The investigation of substrate specificity of individual UDPGTs by expression of cloned genes in COS-7 cells and the use of antibodies will facilitate the identification of enzymes present in perinatal liver.

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Section: Methodsmentioning

confidence: 99%

Development of Human Liver UDP-Glucuronosyltransferases

Burchell¹,

Coughtrie²,

Jackson³

et al. 1989

Dev Pharmacol Ther

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“…Oestrone glucuronyltransferase activity was determined by a modification of the procedure of Rao et al (1976 conducted for 10min at 37°C and terminated by the addition of 1 ml of 10 % (w/v) trichloroacetic acid. The precipitated protein was removed by centrifugation at 10OOg for 10min and a 2ml sample of the clear supernatant was added to I ml of 2.0M-Tris/HCI buffer, pH9.0, containing 3 umol of p-nitrophenol.…”

Section: Methodsmentioning

confidence: 99%

Separation of oestrone UDP-glucuronyltransferase and p-nitrophenol UDP-glucuronyltransferase activities

Tukey¹,

Billings²,

Tephly³

1978

Biochemical Journal

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“…Testosterone, oestrone and ,-oestradiol UDPGT assays were carried out as described by Rao et al. [18,19], except that all steroids were present at 0.6mm. Androsterone UDPGT assay was based on that described by Matsui & Hakozaki [20], except that the steroid was present at 0.1 mm instead of 0.172 mm.…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

“…Androsterone UDPGT assay was based on that described by Matsui & Hakozaki [20], except that the steroid was present at 0.1 mm instead of 0.172 mm. Free steroid and glucuronide fractions were separated as described by Rao et al [18].…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

Multiplicity of UDP-glucuronosyltransferases in fish. Purification and characterization of a phenol UDP-glucuronosyltransferase from the liver of a marine teleost, Pleuronectes platessa

Clarke

George

Burchell

1992

Biochemical Journal

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The aim of this work was to determine if a non-mammalian species had multiple UDP-glucuronosyltransferase (UDPGT) isoforms. At least six highly purified UDPGT isoenzymes were partially resolved by anion-exchange chromatography and UDP-hexanolamine-Sepharose 4B affinity chromatography from liver microsomes of a fish, the plaice. Q-Sepharose FF, chromatofocusing and affinity-chromatographic procedures were employed to separate and purify the phenol UDPGT isoform to apparent homogeneity. The purified enzyme conjugated l-naphthol, but not bilirubin or steroids, and displayed a pl of 7.0 and a subunit molecular mass of 55 kDa. Bilirubin and testosterone UDPGT activities were more labile and, although purified over 200-fold, these preparations also contained the phenol UDPGT and had multiple polypeptides with molecular masses of 52-57 kDa. Antisera to rat bilirubin/phenol UDPGT and testosterone/phenol UDPGT isoforms cross-reacted strongly with the partially purified plaice UDPGT isoforms of molecular masses 52, 53 and 57 kDa and less strongly with phenol UDPGT 54 kDa and 56 kDa isoforms. Fish and mammalian UDPGTs therefore apparently possess a high degree of evolutionary conservation.

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An improved assay for steroid glucuronyltransferase in rat liver microsomes

Cited by 78 publications

References 7 publications

Development of Human Liver UDP-Glucuronosyltransferases

Development of Human Liver UDP-Glucuronosyltransferases

Separation of oestrone UDP-glucuronyltransferase and p-nitrophenol UDP-glucuronyltransferase activities

Multiplicity of UDP-glucuronosyltransferases in fish. Purification and characterization of a phenol UDP-glucuronosyltransferase from the liver of a marine teleost, Pleuronectes platessa

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