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Dimerization of human uridine diphosphate glucuronosyltransferase allozymes 1A1 and 1A9 alters their quercetin glucuronidation activities.

Liu YQ, Yuan LM, Gao ZZ, Xiao YS, Sun HY, Yu LS, Zeng S - Sci Rep (2016)

Bottom Line: Uridine diphosphate glucuronosyltransferase 1A (UGT1A) is a major phase II drug-metabolism enzyme superfamily involved in the glucuronidation of endobiotics and xenobiotics in humans.Many polymorphisms in UGT1A genes are reported to inhibit or decrease UGT1A activity.SNPs of UGT1A altered the ability of protein-protein interaction, resulting in differential FRET efficiencies and donor-acceptor r distances.

View Article: PubMed Central - PubMed

Affiliation: Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Uridine diphosphate glucuronosyltransferase 1A (UGT1A) is a major phase II drug-metabolism enzyme superfamily involved in the glucuronidation of endobiotics and xenobiotics in humans. Many polymorphisms in UGT1A genes are reported to inhibit or decrease UGT1A activity. In this study, two UGT1A1 allozymes, UGT1A1 wild-type and a splice mutant, as well as UGT1A9 wild-type and its three UGT1A9 allozymes, UGT1A9*2(C3Y), UGT1A9*3(M33T), and UGT1A9*5(D256N) were single- or double-expressed in a Bac-to-Bac expression system. Dimerization of UGT1A1 or UGT1A9 allozymes was observed via fluorescence resonance energy transfer (FRET) and co-immunoprecipitation analysis. SNPs of UGT1A altered the ability of protein-protein interaction, resulting in differential FRET efficiencies and donor-acceptor r distances. Dimerization changed the chemical regioselectivity, substrate-binding affinity, and enzymatic activity of UGT1A1 and UGT1A9 in glucuronidation of quercetin. These findings provide molecular insights into the consequences of homozygous and heterozygous UGT1A1 and UGT1A9 allozymes expression on quercetin glucuronidation.

No MeSH data available.


Related in: MedlinePlus

Dimerization of human UGT 1A1 and 1A9 alters their quercetin glucuronidation activities.(A) Domains of UGT1A1*1/*1b and UGT1A9*1/*2/*3/*5; (B) Dimer detection by FRET using CFP and YFP tags; (C) Chemical reaction of quercetin glucuronidation catalyzed by UGT1A.
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f1: Dimerization of human UGT 1A1 and 1A9 alters their quercetin glucuronidation activities.(A) Domains of UGT1A1*1/*1b and UGT1A9*1/*2/*3/*5; (B) Dimer detection by FRET using CFP and YFP tags; (C) Chemical reaction of quercetin glucuronidation catalyzed by UGT1A.

Mentions: In this study (as shown in Fig. 1), UGT1A1 allozymes (UGT1A1*1 and one splice mutant UGT1A1*1b) and UGT1A9 allozymes (UGT1A9*1/*2/*3/*5) were over expressed individually or co-expressed with one other variant in insect cell lines using a Bac-to-Bac expression system, respectively. Homodimeric formation among the wild-type enzyme and variants was determined by quantitative FRET and Co-IP. The enzyme activity and differential chemical regioselectivity were detected by glucuronides formation from quercetin to determine whether the protein-protein interactions between UGT1A allozymes altered the enzymatic characteristics.


Dimerization of human uridine diphosphate glucuronosyltransferase allozymes 1A1 and 1A9 alters their quercetin glucuronidation activities.

Liu YQ, Yuan LM, Gao ZZ, Xiao YS, Sun HY, Yu LS, Zeng S - Sci Rep (2016)

Dimerization of human UGT 1A1 and 1A9 alters their quercetin glucuronidation activities.(A) Domains of UGT1A1*1/*1b and UGT1A9*1/*2/*3/*5; (B) Dimer detection by FRET using CFP and YFP tags; (C) Chemical reaction of quercetin glucuronidation catalyzed by UGT1A.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837415&req=5

f1: Dimerization of human UGT 1A1 and 1A9 alters their quercetin glucuronidation activities.(A) Domains of UGT1A1*1/*1b and UGT1A9*1/*2/*3/*5; (B) Dimer detection by FRET using CFP and YFP tags; (C) Chemical reaction of quercetin glucuronidation catalyzed by UGT1A.
Mentions: In this study (as shown in Fig. 1), UGT1A1 allozymes (UGT1A1*1 and one splice mutant UGT1A1*1b) and UGT1A9 allozymes (UGT1A9*1/*2/*3/*5) were over expressed individually or co-expressed with one other variant in insect cell lines using a Bac-to-Bac expression system, respectively. Homodimeric formation among the wild-type enzyme and variants was determined by quantitative FRET and Co-IP. The enzyme activity and differential chemical regioselectivity were detected by glucuronides formation from quercetin to determine whether the protein-protein interactions between UGT1A allozymes altered the enzymatic characteristics.

Bottom Line: Uridine diphosphate glucuronosyltransferase 1A (UGT1A) is a major phase II drug-metabolism enzyme superfamily involved in the glucuronidation of endobiotics and xenobiotics in humans.Many polymorphisms in UGT1A genes are reported to inhibit or decrease UGT1A activity.SNPs of UGT1A altered the ability of protein-protein interaction, resulting in differential FRET efficiencies and donor-acceptor r distances.

View Article: PubMed Central - PubMed

Affiliation: Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Uridine diphosphate glucuronosyltransferase 1A (UGT1A) is a major phase II drug-metabolism enzyme superfamily involved in the glucuronidation of endobiotics and xenobiotics in humans. Many polymorphisms in UGT1A genes are reported to inhibit or decrease UGT1A activity. In this study, two UGT1A1 allozymes, UGT1A1 wild-type and a splice mutant, as well as UGT1A9 wild-type and its three UGT1A9 allozymes, UGT1A9*2(C3Y), UGT1A9*3(M33T), and UGT1A9*5(D256N) were single- or double-expressed in a Bac-to-Bac expression system. Dimerization of UGT1A1 or UGT1A9 allozymes was observed via fluorescence resonance energy transfer (FRET) and co-immunoprecipitation analysis. SNPs of UGT1A altered the ability of protein-protein interaction, resulting in differential FRET efficiencies and donor-acceptor r distances. Dimerization changed the chemical regioselectivity, substrate-binding affinity, and enzymatic activity of UGT1A1 and UGT1A9 in glucuronidation of quercetin. These findings provide molecular insights into the consequences of homozygous and heterozygous UGT1A1 and UGT1A9 allozymes expression on quercetin glucuronidation.

No MeSH data available.


Related in: MedlinePlus