Purification and properties of microsomal UDP-glucuronosyltransferase from rat liver.

Gorski, Jeffrey P.; Kasper, Charles B.

doi:10.1016/s0021-9258(17)40661-2

Cited by 101 publications

(8 citation statements)

References 48 publications

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“…Male Sprague-Dawley rats (150-175 g) were injected with sodium phenobarbital, 0.12 g/kg of body weight, according to Khandwala & Kasper (1973). Gradient-purified microsomal membrane was prepared from excised liver and stored frozen as previously reported (Gorski & Kasper, 1977).…”

Section: Methodsmentioning

confidence: 99%

“…Delipidated microsomal protein (approximately 200 pg in 0.1 mL) was added to 0.2 mL of liposomes containing 3 Mmol of phospholipid and the mixture vortexed intermittently for 10 min at room temperature. Approximately 150 Mg of reconstituted enzyme was used per assay (Gorski & Kasper, 1977). Deoxycholate solubilized membrane was assayed by the same procedure after sixfold dilution with 20 mM Tris-acetate, pH 7.5, containing 0.10-0.12% (w/v) sodium deoxycholate, 120 mM sodium chloride, and 0.02% sodium azide.…”

Section: Methodsmentioning

confidence: 99%

“…Kinetic Assay of Reconstituted Transferase. Delipidated microsomal protein (approximately 300 pg in 0.15 mL) was mixed with an equal volume of liposomes (0.5-0.85 Mmol of phospholipids) in a cuvette and assayed as previously described (Gorski & Kasper, 1977). p-Nitrophenol displays a broad absorbance spectrum centered at 400 nm, and operationally a wavelength was chosen so as to obtain an initial absorbance between 1.5 and 2.0 OD.…”

Section: Methodsmentioning

confidence: 99%

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UDP-glucuronosyltransferase: phospholipid dependence and properties of the reconstituted apoenzyme

Gorski¹,

Kasper²

1978

Biochemistry

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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UDP-glucuronosyltransferase: phospholipid dependence and properties of the reconstituted apoenzyme

Gorski¹,

Kasper²

1978

Biochemistry

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“…Rat hepatic UDP-glucuronyltransferase with activity towards phenolic substrates has been purified to apparent homogeneity in this (Burchell, 1977(Burchell, , 1978) and other laboratories (Gorski & Kasper, 1977;Bock et al, 1979). Transferase activity towards testosterone, when looked for, was either not detectable or extremely low (Burchell, 1978;Bock et al, 1979).…”

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confidence: 88%

The separation and purification of rat liver UDP-glucuronyltransferase activities towards testosterone and oestrone

Weatherill¹,

Burchell²

1980

Biochemical Journal

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1. Reconstitution of UDP-glucuronyltransferase preparations with phosphataidylcholine liposomes facilitated the purification of testosterone UDP-glucuronyltransferase. 2. Transferase activity towards testosterone co-purifies with that towards 4-nitrophenol. 3. UDP-glucuronyltransferase activity towards oestrone was separated from that towards testosterone. 4. These results suggest that testosterone and 4-nitrophenol may be glucuronidated by a different form of UDP-glucuronyltransferase from the one glucuronidating oestrone.

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“…From our point of view, three major breakthroughs occurred which allowed for improvements in UDPGT purification. Gorski and Kasper (25) used a UDP-hexanolamine Sepharose 4B affinity procedure which facilitated the purification of a rat liver microsomal UDPGT reacting with p-nitrophenol. This procedure was useful but did not allow for separation of UDPGTs if several UDPGTs were present when applied to the resin.…”

Section: Introductionmentioning

confidence: 99%

Isolation and purification of UDP-glucuronosyltransferases

Tephly¹

1990

Chem. Res. Toxicol.

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UDPGTs are members of a class of enzymes located in the endoplasmic reticulum and are encoded by a multigene family. These proteins are responsible for the glucuronidation of hundreds of xenobiotics of many chemical classes and many endogenous substances such as steroid hormones, bile acids, and bilirubin. There are a number of UDPGTs which have been identified by purification and characterization studies and a significant number which have been characterized by expression of cDNAs. On the basis of the primary structures elucidated they appear to have marked similarities (5) and are highly conserved. However, key differences in their functional properties appear to depend primarily on differences in amino acid sequences at or about the NH2-terminal area of the protein (5). Many of the UDPGTs have an extraordinarily broad substrate specificity; a few, however, are relatively specific for a given class of substrate (morphine, DT-1 UDPGTs). This places a burden on investigators to clearly identify which substrate and how many UDPGTs will be involved in any analysis of rates of glucuronidation in microsomal preparations. Caution should also be advised for extrapolation of data from hepatic microsomes of experimental animals to human hepatic microsomal preparations because human liver microsomes possess UDPGTs which are qualitatively different and, in certain cases, UDPGTs are present in human liver which are not present in lower animals.

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Purification and properties of microsomal UDP-glucuronosyltransferase from rat liver.

Cited by 101 publications

References 48 publications

UDP-glucuronosyltransferase: phospholipid dependence and properties of the reconstituted apoenzyme

UDP-glucuronosyltransferase: phospholipid dependence and properties of the reconstituted apoenzyme

The separation and purification of rat liver UDP-glucuronyltransferase activities towards testosterone and oestrone

Isolation and purification of UDP-glucuronosyltransferases

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