Functional analysis of UGT1A4 P24T and UGT1A4 L48V variant enzymes

Jin Zhou, Upendra A. Argikar, Rory P Remmel

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Aim: To investigate the effects of two nonsynonymous SNPs, UGT1A4*2 (rs: 6755571, 70C>A, P24T) and UGT1A4* (rs: 2011425, 142T>G, L48V), on the function of UGT1A4 against dihydrotestosterone (DHT), transandrosterone (t-AND), lamotrigine (LTG) and tamoxifen (TAM). Materials & methods: Detailed kinetic experiments were conducted with recombinant UGT1A4 wild-type, UGT1A4 P24T and UGT1A4 L48V, which were overexpressed in HEK293 cell lines. The kinetic profiles and kinetic parameters (K m, V max and CL int) obtained with either UGT1A4 P24T or UGT1A4 L48V were compared with those obtained with the wild-type enzyme. The interaction of TAM on UG1A4-catalyzed DHT glucuronidation was also investigated with the three UGT1A4 polymorphic enzymes. Results: UGT1A4 L48V had higher enzyme efficiency (CL int) compared with wild-type UGT1A4 on DHT glucuronidation; UGT1A4 P24T and UGT1A4 L48V had lower CL int than wild-type UGT1A4 for t-AND and LTG glucuronidation. The TAM CL int with UGT1A4 P24T and UGT1A4 L48V glucuronidation and the UGT1A4 P24T-catalyzed DHT glucuronidation were, on the other hand, similar to those of the wild-type enzyme. With all three enzymes, TAM activated UGT1A4-catalyzed DHT glucuronidation in a concentration-dependent fashion. Conclusion: Decreased CL int of UGT1A4 P24T and UGT1A4 L48V on LTG glucuronidation may lead to interindividual variations in LTG metabolism in vivo. However, it is less likely that these polymorphisms would have impact on DHT and t-AND metabolism in vivo because these compounds are glucuronidated by multiple enzymes.

Original languageEnglish (US)
Pages (from-to)1671-1679
Number of pages9
JournalPharmacogenomics
Volume12
Issue number12
DOIs
StatePublished - Dec 1 2011

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Dihydrotestosterone
Tamoxifen
Enzymes
bilirubin glucuronoside glucuronosyltransferase
HEK293 Cells
Single Nucleotide Polymorphism
Cell Line
lamotrigine

Keywords

  • UGT1A4
  • dihydrotestosterone
  • enzyme kinetic
  • glucuronidation
  • heteroactivation
  • lamotrigine
  • polymorphism
  • tamoxifen
  • transandrosterone

Cite this

Functional analysis of UGT1A4 P24T and UGT1A4 L48V variant enzymes. / Zhou, Jin; Argikar, Upendra A.; Remmel, Rory P.

In: Pharmacogenomics, Vol. 12, No. 12, 01.12.2011, p. 1671-1679.

Research output: Contribution to journalArticle

Zhou, Jin ; Argikar, Upendra A. ; Remmel, Rory P. / Functional analysis of UGT1A4 P24T and UGT1A4 L48V variant enzymes. In: Pharmacogenomics. 2011 ; Vol. 12, No. 12. pp. 1671-1679.
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abstract = "Aim: To investigate the effects of two nonsynonymous SNPs, UGT1A4*2 (rs: 6755571, 70C>A, P24T) and UGT1A4* (rs: 2011425, 142T>G, L48V), on the function of UGT1A4 against dihydrotestosterone (DHT), transandrosterone (t-AND), lamotrigine (LTG) and tamoxifen (TAM). Materials & methods: Detailed kinetic experiments were conducted with recombinant UGT1A4 wild-type, UGT1A4 P24T and UGT1A4 L48V, which were overexpressed in HEK293 cell lines. The kinetic profiles and kinetic parameters (K m, V max and CL int) obtained with either UGT1A4 P24T or UGT1A4 L48V were compared with those obtained with the wild-type enzyme. The interaction of TAM on UG1A4-catalyzed DHT glucuronidation was also investigated with the three UGT1A4 polymorphic enzymes. Results: UGT1A4 L48V had higher enzyme efficiency (CL int) compared with wild-type UGT1A4 on DHT glucuronidation; UGT1A4 P24T and UGT1A4 L48V had lower CL int than wild-type UGT1A4 for t-AND and LTG glucuronidation. The TAM CL int with UGT1A4 P24T and UGT1A4 L48V glucuronidation and the UGT1A4 P24T-catalyzed DHT glucuronidation were, on the other hand, similar to those of the wild-type enzyme. With all three enzymes, TAM activated UGT1A4-catalyzed DHT glucuronidation in a concentration-dependent fashion. Conclusion: Decreased CL int of UGT1A4 P24T and UGT1A4 L48V on LTG glucuronidation may lead to interindividual variations in LTG metabolism in vivo. However, it is less likely that these polymorphisms would have impact on DHT and t-AND metabolism in vivo because these compounds are glucuronidated by multiple enzymes.",
keywords = "UGT1A4, dihydrotestosterone, enzyme kinetic, glucuronidation, heteroactivation, lamotrigine, polymorphism, tamoxifen, transandrosterone",
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AU - Argikar, Upendra A.

AU - Remmel, Rory P

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N2 - Aim: To investigate the effects of two nonsynonymous SNPs, UGT1A4*2 (rs: 6755571, 70C>A, P24T) and UGT1A4* (rs: 2011425, 142T>G, L48V), on the function of UGT1A4 against dihydrotestosterone (DHT), transandrosterone (t-AND), lamotrigine (LTG) and tamoxifen (TAM). Materials & methods: Detailed kinetic experiments were conducted with recombinant UGT1A4 wild-type, UGT1A4 P24T and UGT1A4 L48V, which were overexpressed in HEK293 cell lines. The kinetic profiles and kinetic parameters (K m, V max and CL int) obtained with either UGT1A4 P24T or UGT1A4 L48V were compared with those obtained with the wild-type enzyme. The interaction of TAM on UG1A4-catalyzed DHT glucuronidation was also investigated with the three UGT1A4 polymorphic enzymes. Results: UGT1A4 L48V had higher enzyme efficiency (CL int) compared with wild-type UGT1A4 on DHT glucuronidation; UGT1A4 P24T and UGT1A4 L48V had lower CL int than wild-type UGT1A4 for t-AND and LTG glucuronidation. The TAM CL int with UGT1A4 P24T and UGT1A4 L48V glucuronidation and the UGT1A4 P24T-catalyzed DHT glucuronidation were, on the other hand, similar to those of the wild-type enzyme. With all three enzymes, TAM activated UGT1A4-catalyzed DHT glucuronidation in a concentration-dependent fashion. Conclusion: Decreased CL int of UGT1A4 P24T and UGT1A4 L48V on LTG glucuronidation may lead to interindividual variations in LTG metabolism in vivo. However, it is less likely that these polymorphisms would have impact on DHT and t-AND metabolism in vivo because these compounds are glucuronidated by multiple enzymes.

AB - Aim: To investigate the effects of two nonsynonymous SNPs, UGT1A4*2 (rs: 6755571, 70C>A, P24T) and UGT1A4* (rs: 2011425, 142T>G, L48V), on the function of UGT1A4 against dihydrotestosterone (DHT), transandrosterone (t-AND), lamotrigine (LTG) and tamoxifen (TAM). Materials & methods: Detailed kinetic experiments were conducted with recombinant UGT1A4 wild-type, UGT1A4 P24T and UGT1A4 L48V, which were overexpressed in HEK293 cell lines. The kinetic profiles and kinetic parameters (K m, V max and CL int) obtained with either UGT1A4 P24T or UGT1A4 L48V were compared with those obtained with the wild-type enzyme. The interaction of TAM on UG1A4-catalyzed DHT glucuronidation was also investigated with the three UGT1A4 polymorphic enzymes. Results: UGT1A4 L48V had higher enzyme efficiency (CL int) compared with wild-type UGT1A4 on DHT glucuronidation; UGT1A4 P24T and UGT1A4 L48V had lower CL int than wild-type UGT1A4 for t-AND and LTG glucuronidation. The TAM CL int with UGT1A4 P24T and UGT1A4 L48V glucuronidation and the UGT1A4 P24T-catalyzed DHT glucuronidation were, on the other hand, similar to those of the wild-type enzyme. With all three enzymes, TAM activated UGT1A4-catalyzed DHT glucuronidation in a concentration-dependent fashion. Conclusion: Decreased CL int of UGT1A4 P24T and UGT1A4 L48V on LTG glucuronidation may lead to interindividual variations in LTG metabolism in vivo. However, it is less likely that these polymorphisms would have impact on DHT and t-AND metabolism in vivo because these compounds are glucuronidated by multiple enzymes.

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KW - dihydrotestosterone

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KW - heteroactivation

KW - lamotrigine

KW - polymorphism

KW - tamoxifen

KW - transandrosterone

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