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Identifying and applying a highly selective probe to simultaneously determine the O-glucuronidation activity of human UGT1A3 and UGT1A4.

Jiang L, Liang SC, Wang C, Ge GB, Huo XK, Qi XY, Deng S, Liu KX, Ma XC - Sci Rep (2015)

Bottom Line: Glucuronidation mediated by uridine 5'-diphospho (UDP)-glucuronosyltransferase is an important detoxification pathway.However, identifying a selective probe of UDP- glucuronosyltransferase is complicated because of the significant overlapping substrate specificity displayed by the enzyme.DACB was well characterized as a probe for simultaneously determining the catalytic activities of O-glucuronidation mediated by UGT1A3 and UGT1A4 from various enzyme sources, through a sensitive analysis method.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Key Laboratory of Pharmacokinetic and Drug Transport of Liaoning, Academy of Integrative Medicine, Dalian Medical University, Dalian, 116044, China.

ABSTRACT
Glucuronidation mediated by uridine 5'-diphospho (UDP)-glucuronosyltransferase is an important detoxification pathway. However, identifying a selective probe of UDP- glucuronosyltransferase is complicated because of the significant overlapping substrate specificity displayed by the enzyme. In this paper, desacetylcinobufagin (DACB) 3-O- and 16-O-glucuronidation were found to be isoform-specific probe reactions for UGT1A4 and UGT1A3, respectively. DACB was well characterized as a probe for simultaneously determining the catalytic activities of O-glucuronidation mediated by UGT1A3 and UGT1A4 from various enzyme sources, through a sensitive analysis method.

No MeSH data available.


Isoform specificity of 3β- (A) and 16β-O-glucuronidation (B).
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f2: Isoform specificity of 3β- (A) and 16β-O-glucuronidation (B).

Mentions: DACB glucuronidation by recombinant UGT Supersomes™ was investigated using a panel of fourteen recombinant UGT isozymes (1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B10, 2B11, 2B15 and 2B17). Surprisingly, UGT1A4 mainly dominated the formation of M-1 at three different substrate concentrations (6, 60 and 600 μM) with a high selectivity (Fig. 2A). Additionally, Fig. 2B indicated that UGT1A3 had a good selectivity in promoting M-2 formation. Only UGT1A1 displayed the limited ability to generate M-2 at the highest substrate concentration (600 μM), which was 30 times less than UGT1A3 (Fig. S5). Up to now, DACB is the most selective probe for O-glucuronidation simultaneously mediated by UGT1A3 and UGT1A4.


Identifying and applying a highly selective probe to simultaneously determine the O-glucuronidation activity of human UGT1A3 and UGT1A4.

Jiang L, Liang SC, Wang C, Ge GB, Huo XK, Qi XY, Deng S, Liu KX, Ma XC - Sci Rep (2015)

Isoform specificity of 3β- (A) and 16β-O-glucuronidation (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401096&req=5

f2: Isoform specificity of 3β- (A) and 16β-O-glucuronidation (B).
Mentions: DACB glucuronidation by recombinant UGT Supersomes™ was investigated using a panel of fourteen recombinant UGT isozymes (1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B10, 2B11, 2B15 and 2B17). Surprisingly, UGT1A4 mainly dominated the formation of M-1 at three different substrate concentrations (6, 60 and 600 μM) with a high selectivity (Fig. 2A). Additionally, Fig. 2B indicated that UGT1A3 had a good selectivity in promoting M-2 formation. Only UGT1A1 displayed the limited ability to generate M-2 at the highest substrate concentration (600 μM), which was 30 times less than UGT1A3 (Fig. S5). Up to now, DACB is the most selective probe for O-glucuronidation simultaneously mediated by UGT1A3 and UGT1A4.

Bottom Line: Glucuronidation mediated by uridine 5'-diphospho (UDP)-glucuronosyltransferase is an important detoxification pathway.However, identifying a selective probe of UDP- glucuronosyltransferase is complicated because of the significant overlapping substrate specificity displayed by the enzyme.DACB was well characterized as a probe for simultaneously determining the catalytic activities of O-glucuronidation mediated by UGT1A3 and UGT1A4 from various enzyme sources, through a sensitive analysis method.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Key Laboratory of Pharmacokinetic and Drug Transport of Liaoning, Academy of Integrative Medicine, Dalian Medical University, Dalian, 116044, China.

ABSTRACT
Glucuronidation mediated by uridine 5'-diphospho (UDP)-glucuronosyltransferase is an important detoxification pathway. However, identifying a selective probe of UDP- glucuronosyltransferase is complicated because of the significant overlapping substrate specificity displayed by the enzyme. In this paper, desacetylcinobufagin (DACB) 3-O- and 16-O-glucuronidation were found to be isoform-specific probe reactions for UGT1A4 and UGT1A3, respectively. DACB was well characterized as a probe for simultaneously determining the catalytic activities of O-glucuronidation mediated by UGT1A3 and UGT1A4 from various enzyme sources, through a sensitive analysis method.

No MeSH data available.