Author/Authors :
Wada، نويسنده , , Keiko and Takeuchi، نويسنده , , Atsuko and Saiki، نويسنده , , Kayoko and Sutomo، نويسنده , , Retno and Van Rostenberghe، نويسنده , , Hans and Yusoff، نويسنده , , Narazah Mohd and Laosombat، نويسنده , , Vichai and Sadewa، نويسنده , , Ahmad Hamim and Talib، نويسنده , , Norlelawati Abdul and Yusoff، نويسنده , , Surini and Lee، نويسنده , , Myeong Jin and Ayaki، نويسنده , , Hitoshi a، نويسنده ,
Abstract :
Mutations in the gene encoding UDP-glucuronosyltransferase 1A1 (UGT1A1) may reduce the glucuronidation of estradiol, bilirubin, etc. In the present study, we used a liquid chromatography–tandem mass spectrometry (LC/MS/MS) method to assay the activities of recombinant mutated UGT1A1 toward 17β-estradiol (E2), by determining its glucuronide (E2G) content. Direct evidence for glucuronide formation was provided by E2G-specific ion peaks. The UGT1A1 activities of G71R (exon 1), F83L (exon 1), I322V (exon 2) and G493R (exon 5) mutants were 24, 30, 18 and 0.6% of the normal UGT1A1 activity, respectively. In conclusion, our study showed that LC/MS/MS enabled accurate evaluation of the effects of mutations on recombinant UGT1A1 activity towards E2.
