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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.


The correlation analysis between the expression of UGT isoforms and the DACB glucuronidation rate in individual HLMs.(A) Western blots of UGT1A3 in individual HLMs; (B) the correlation between UGT1A3 expression and DACB 16-O-glucuronidation rates in 12 individual HLMs; (C) Western blots of UGT1A4 in individual HLMs; (D) the correlation between UGT1A4 expression and DACB 3-O-glucuronidation rates in 12 individual HLMs.
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f5: The correlation analysis between the expression of UGT isoforms and the DACB glucuronidation rate in individual HLMs.(A) Western blots of UGT1A3 in individual HLMs; (B) the correlation between UGT1A3 expression and DACB 16-O-glucuronidation rates in 12 individual HLMs; (C) Western blots of UGT1A4 in individual HLMs; (D) the correlation between UGT1A4 expression and DACB 3-O-glucuronidation rates in 12 individual HLMs.

Mentions: To evaluate the metabolic activities of UGT1A3 and UGT1A4 in HLMs accurately, the glucuronidation activities of UGT1A3 and UGT1A4 in a panel of HLMs from 12 individuals were determined using the formation of M-1 and M-2, respectively. Additionally, the expression levels of UGT1A3 and UGT1A4 in these individual HLMs were measured using western blot technique. The correlation analysis of DACB 3-O- and 16-O-glucuronidation rates with the expression levels of UGT1A3 and UGT1A4 in the corresponding individual HLMs, showed strong correlations with the correlation coefficients (R) in the range of 0.79 to 0.82 (P < 0.05) (Fig. 5). Our results demonstrated that DACB is a highly selective probe to simultaneously measure the O-glucuronidation bioactivities of UGT1A3 and UGT1A4 in various biological samples.


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)

The correlation analysis between the expression of UGT isoforms and the DACB glucuronidation rate in individual HLMs.(A) Western blots of UGT1A3 in individual HLMs; (B) the correlation between UGT1A3 expression and DACB 16-O-glucuronidation rates in 12 individual HLMs; (C) Western blots of UGT1A4 in individual HLMs; (D) the correlation between UGT1A4 expression and DACB 3-O-glucuronidation rates in 12 individual HLMs.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: The correlation analysis between the expression of UGT isoforms and the DACB glucuronidation rate in individual HLMs.(A) Western blots of UGT1A3 in individual HLMs; (B) the correlation between UGT1A3 expression and DACB 16-O-glucuronidation rates in 12 individual HLMs; (C) Western blots of UGT1A4 in individual HLMs; (D) the correlation between UGT1A4 expression and DACB 3-O-glucuronidation rates in 12 individual HLMs.
Mentions: To evaluate the metabolic activities of UGT1A3 and UGT1A4 in HLMs accurately, the glucuronidation activities of UGT1A3 and UGT1A4 in a panel of HLMs from 12 individuals were determined using the formation of M-1 and M-2, respectively. Additionally, the expression levels of UGT1A3 and UGT1A4 in these individual HLMs were measured using western blot technique. The correlation analysis of DACB 3-O- and 16-O-glucuronidation rates with the expression levels of UGT1A3 and UGT1A4 in the corresponding individual HLMs, showed strong correlations with the correlation coefficients (R) in the range of 0.79 to 0.82 (P < 0.05) (Fig. 5). Our results demonstrated that DACB is a highly selective probe to simultaneously measure the O-glucuronidation bioactivities of UGT1A3 and UGT1A4 in various biological samples.

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.