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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.


DACB 3β- and 16β-O-glucuronidation by UGT1A4 and UGT1A3, respectively.
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f1: DACB 3β- and 16β-O-glucuronidation by UGT1A4 and UGT1A3, respectively.

Mentions: After screening a series of natural and transformed bufadienolides (Figure S1) using human UGT isoforms in the present paper, we found an isoform-specific probe substrate (desacetylcinobufagin, DACB, Figure 1) for simultaneously determining the O-glucuronidation activities of UGT1A3 and UGT1A4 in the different enzyme resources. The selectivities for UGTs 1A3 and 1A4 were determined by chemical inhibition testings, screening assays with human UGT isoforms, and the correlation assays. Our results indicated that UGT1A4 catalyzed 3-O-glucuronidation of DACB with excellent selectivity, and UGT1A3 was found to dominantly catalyze the 16-O-glucuronidation of DACB, based on the kinetic studies of HLMs and UGT isoforms. It is firstly reported the simultaneous and specific determination of O- glucuronidation activities of 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)

DACB 3β- and 16β-O-glucuronidation by UGT1A4 and UGT1A3, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: DACB 3β- and 16β-O-glucuronidation by UGT1A4 and UGT1A3, respectively.
Mentions: After screening a series of natural and transformed bufadienolides (Figure S1) using human UGT isoforms in the present paper, we found an isoform-specific probe substrate (desacetylcinobufagin, DACB, Figure 1) for simultaneously determining the O-glucuronidation activities of UGT1A3 and UGT1A4 in the different enzyme resources. The selectivities for UGTs 1A3 and 1A4 were determined by chemical inhibition testings, screening assays with human UGT isoforms, and the correlation assays. Our results indicated that UGT1A4 catalyzed 3-O-glucuronidation of DACB with excellent selectivity, and UGT1A3 was found to dominantly catalyze the 16-O-glucuronidation of DACB, based on the kinetic studies of HLMs and UGT isoforms. It is firstly reported the simultaneous and specific determination of O- glucuronidation activities of 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.