Molecular cloning and functional analysis of minipig UDP-glucuronosyltransferase 1A6. (3rd March 2016)

• Record Type: Journal Article
• Authors: Miyake, Yuuka; Hirose, Riho; Isobe, Takashi; Hanioka, Nobumitsu
• Is Part Of: Xenobiotica. Volume 46:Number 3(2016:Mar.)
• Page Start: 193
• Page End: 199
• Publication Date: 2016-03-03
• Subjects: Glucuronidation -- minipig -- serotonin -- UDP-glucuronosyltransferase -- UGT1A6; Metabolism -- Periodicals; Drugs -- Physiological effect -- Periodicals; Food additives -- Periodicals; Chemicals -- Physiological effect -- Periodicals; Biochemistry -- Periodicals; Pharmaceutical Preparations -- metabolism -- Periodicals; Metabolism -- Periodicals; 574.133
• DOI: 10.3109/00498254.2015.1060373
• Languages: English
• ISSNs: 0049-8254

Abstract

Abstract: 1. UDP-glucuronosyltransferase 1A6 (UGT1A6) plays important roles in the glucuronidation of numerous drugs, environmental pollutants, and endogenous substances. Minipigs have been used as experimental animals in pharmacological and toxicological studies because many of their physiological characteristics are similar to those of humans. The aim of the present study was to examine similarities and differences in the enzymatic properties of UGT1A6 between humans and minipigs. 2. Minipig UGT1A6 (mpUGT1A6) cDNA was cloned by the RACE method, and the corresponding proteins were expressed in insect cells. The enzymatic function of mpUGT1A6 was analyzed by the kinetics of serotonin glucuronidation. 3. Amino acid homology between human UGT1A6 (hUGT1A6) and mpUGT1A6 was 79.9%. The kinetics of serotonin glucuronidation by recombinant hUGT1A6 and mpUGT1A6 enzymes fit the Michaelis–Menten equation. The K m, V max, and CL int values of hUGT1A6 were 10.5 mM, 4.04 nmol/min/mg protein, and 0.39 µL/min/mg protein, respectively. The K m value of mpUGT1A6 was similar to that of hUGT1A6, whereas the V max and CL int values of mpUGT1A6 were approximately 2-fold higher than those of hUGT1A6. 4. These results suggest that the enzymatic properties of UGT1A6 enzymes are moderately different between humans and minipigs.