Evaluation of variation of glucuronidation activity and substrate specificity caused by difference in the proportion of expression level of UGT isozimes in living cells

Research Project

Project/Area Number	24790139
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Medical pharmacy
Research Institution	Hokkaido University
Principal Investigator	TAKEKUMA Yoh 北海道大学, 薬学研究科(研究院), 准教授 (00396293)
Project Period (FY)	2012-04-01 – 2014-03-31
Project Status	Completed (Fiscal Year 2013)
Budget Amount *help	¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000) Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000) Fiscal Year 2012: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords	UGT / 生細胞 / グルクロン酸抱合 / 薬物代謝 / グルクロン酸転移酵素
Research Abstract	The aim of this study was to clarify the factor of gap between UGT activity evaluated by in vitro and in vivo assay. Then we have developed an assay method to determine UGT activity in living cells (as near in vivo model). The method is as follows: LLC-PK1 cells were incubated with substrate for 2 hours on ice. After incubating, the cells were incubated on water bath at 37 degree Celsius for enzymes and transporters working. And then metabolites were extracellularly excreted and quantified. Furthermore, we established stable HeLa cell line expressing UGT1A9 and introduced UGT2B4 into it. UGT1A9 activity of HeLa cells expressing UGT2B4 reduced. This result indicated the protein-protein interaction between the UGTs.

Report

(3 results)