Human UDP-glucuronosyltransferase UGT2A2: cDNA construction, expression, and functional characterization in comparison with UGT2A1 and UGT2A3.

ABSTRACT:

Objectives

Characterize the expression and glucuronidation activities of the human uridine 5'-diphospho (UDP)-glucuronosyltransferase (UGT) 2A2.

Method

UGT2A1 was cloned from nasal mucosa mRNA. Synthetic cDNA for UGT2A2 was constructed assuming exon sharing between UGT2A1 and UGT2A2 (Mackenzie et al., Pharmacogenetics and Genomics 2005, 15:677-685). Exon 1 of UGT2A2 was amplified from genomic DNA and combined with exons 2-6 of UGT2A1. UGT2A3 was cloned from liver mRNA. Quantitative reverse-transcribed-PCR (RT-PCR) was used to evaluate the expression of all the three UGTs of subfamily 2A in different tissues. Recombinant UGT2A1, UGT2A2 and UGT2A3 were expressed in baculovirus-infected insect cells and analyzed for glucuronidation activity towards different substrates.

Resu

SUBMITTER: Sneitz N 

PROVIDER: S-EPMC2928392 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature