Molecular Cloning of the Baboon UDP-glucuronosyltransferase 2B Gene Family and Their Activity in Conjugating Morphine

Overview

Journal Drug Metab Dispos

Specialty Pharmacology

Date 2010 Jan 15

PMID 20071451

Citations 8

Authors

Kirsten Abildskov

Piper Weldy

Marianne Garland

Affiliations

Soon will be listed here.

Abstract

Glucuronidation by UDP-glucuronyltransferase 2B enzymes (UGT2Bs) is a major pathway for the elimination of endobiotics and xenobiotics, including therapeutic drugs. Morphine, a probe drug for UGT2B7, is metabolized to morphine-3-beta-glucuronide (M3G) and morphine-6-beta-glucuronide (M6G) in humans. Morphine has been used in a series of experiments in the baboon to characterize developmental changes in fetal glucuronidation. This study identifies the baboon UGT2B family of enzymes, compares them with that of the human and the monkey (Macaca fascicularis), and measures the activity of the individual baboon UGT2Bs toward morphine. UGT2B cDNAs were cloned from the liver of adult and newborn baboons and expressed in human embryonic kidney 293 cells. The UGT activity toward morphine was assessed by the rate of formation of M3G and M6G by high-performance liquid chromatography. Eight baboon UGT2Bs were cloned and identified: UGT2B41 and UGT2B42, which are 90% homologous to human UGT2B4; UGT2B43, which is 93% homologous to human UGT2B15; and UGT2B39, UGT2B40, UGT2B44, UGT2B45, and UGT2B46, which are 89 to 91% homologous to human UGT2B7. Homology between baboon and monkey UGT2B ranged from 92.6 to 99.1%, with the primary protein structure of UGT2B43 being 99.1% identical to monkey UGT2B20, including a unique R96I substitution. Gene conversion interfered with the phylogenetic signal in the baboon UGT2B7-like and the monkey UGT2B4-like groups and led to concerted evolution of these enzymes. All of the baboon UGT2Bs metabolized morphine to both M3G and M6G. This study lays the foundation for investigating the regulation of UGT2B enzymes during fetal and neonatal development in the baboon.

Citing Articles

Histomorphological and ultrastructural cadmium-induced kidney injuries and precancerous lesions in rats and screening for biomarkers.

Wan X, Xing Z, Ouyang J, Liu H, Cheng C, Luo T Biosci Rep. 2022; 42(6).

PMID: 35678542 PMC: 9202506. DOI: 10.1042/BSR20212516.

Alterations of Cytochrome P450s and UDP-Glucuronosyltransferases in Brain Under Diseases and Their Clinical Significances.

Sheng Y, Yang H, Wu T, Zhu L, Liu L, Liu X Front Pharmacol. 2021; 12:650027.

PMID: 33967789 PMC: 8097730. DOI: 10.3389/fphar.2021.650027.

Predicting and Understanding the Human Microbiome's Impact on Pharmacology.

Hitchings R, Kelly L Trends Pharmacol Sci. 2019; 40(7):495-505.

PMID: 31171383 PMC: 6758919. DOI: 10.1016/j.tips.2019.04.014.

Nanocrystalization: An Emerging Technology to Enhance the Bioavailability of Poorly Soluble Drugs.

Joshi K, Chandra A, Jain K, Talegaonkar S Pharm Nanotechnol. 2019; 7(4):259-278.

PMID: 30961518 PMC: 6967137. DOI: 10.2174/2211738507666190405182524.

siRNA capsulated brain-targeted nanoparticles specifically knock down OATP2B1 in mice: a mechanism for acute morphine tolerance suppression.

Yang Z, Li L, Wang L, Xu M, An S, Jiang C Sci Rep. 2016; 6:33338.

PMID: 27629937 PMC: 5024137. DOI: 10.1038/srep33338.

References

Posada D, Crandall K, Holmes E . Recombination in evolutionary genomics. Annu Rev Genet. 2002; 36:75-97. DOI: 10.1146/annurev.genet.36.040202.111115. View

Green M, King C, Mojarrabi B, Mackenzie P, Tephly T . Glucuronidation of amines and other xenobiotics catalyzed by expressed human UDP-glucuronosyltransferase 1A3. Drug Metab Dispos. 1998; 26(6):507-12. View

Coffman B, Kearney W, Goldsmith S, Knosp B, Tephly T . Opioids bind to the amino acids 84 to 118 of UDP-glucuronosyltransferase UGT2B7. Mol Pharmacol. 2003; 63(2):283-8. DOI: 10.1124/mol.63.2.283. View

Mackenzie P, Bock K, Burchell B, Guillemette C, Ikushiro S, Iyanagi T . Nomenclature update for the mammalian UDP glycosyltransferase (UGT) gene superfamily. Pharmacogenet Genomics. 2005; 15(10):677-85. DOI: 10.1097/01.fpc.0000173483.13689.56. View

DROUIN G, Prat F, Ell M, Clarke G . Detecting and characterizing gene conversions between multigene family members. Mol Biol Evol. 1999; 16(10):1369-90. DOI: 10.1093/oxfordjournals.molbev.a026047. View

Cox L, Mahaney M, VandeBerg J, Rogers J . A second-generation genetic linkage map of the baboon (Papio hamadryas) genome. Genomics. 2006; 88(3):274-81. DOI: 10.1016/j.ygeno.2006.03.020. View

. Initial sequence of the chimpanzee genome and comparison with the human genome. Nature. 2005; 437(7055):69-87. DOI: 10.1038/nature04072. View

Garland M, Szeto H, Daniel S, Tropper P, Myers M, Stark R . Placental transfer and fetal metabolism of zidovudine in the baboon. Pediatr Res. 1998; 44(1):47-53. DOI: 10.1203/00006450-199807000-00008. View

Nebert D, Dieter M . The evolution of drug metabolism. Pharmacology. 2000; 61(3):124-35. DOI: 10.1159/000028393. View

10.

Karere G, Froenicke L, Millon L, Womack J, Lyons L . A high-resolution radiation hybrid map of rhesus macaque chromosome 5 identifies rearrangements in the genome assembly. Genomics. 2008; 92(4):210-8. PMC: 2605074. DOI: 10.1016/j.ygeno.2008.05.013. View

11.

Menard V, Eap O, Harvey M, Guillemette C, Levesque E . Copy-number variations (CNVs) of the human sex steroid metabolizing genes UGT2B17 and UGT2B28 and their associations with a UGT2B15 functional polymorphism. Hum Mutat. 2009; 30(9):1310-9. DOI: 10.1002/humu.21054. View

12.

Mackenzie P, Owens I, Burchell B, Bock K, Bairoch A, Belanger A . The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. Pharmacogenetics. 1997; 7(4):255-69. DOI: 10.1097/00008571-199708000-00001. View

13.

Sawyer S . Statistical tests for detecting gene conversion. Mol Biol Evol. 1989; 6(5):526-38. DOI: 10.1093/oxfordjournals.molbev.a040567. View

14.

Beaulieu M, Levesque E, Barbier O, Turgeon D, Belanger G, Hum D . Isolation and characterization of a simian UDP-glucuronosyltransferase UGT2B18 active on 3-hydroxyandrogens. J Mol Biol. 1998; 275(5):785-94. DOI: 10.1006/jmbi.1997.1486. View

15.

Garland M, Abildskov K, Taylor S, Benzeroual K, Caspersen C, Arroyo S . Fetal morphine metabolism and clearance are constant during late gestation. Drug Metab Dispos. 2006; 34(4):636-46. DOI: 10.1124/dmd.105.007567. View

16.

Belanger G, Barbier O, Hum D, Belanger A . Molecular cloning, expression and characterization of a monkey steroid UDP-glucuronosyltransferase, UGT2B19, that conjugates testosterone. Eur J Biochem. 1999; 260(3):701-8. DOI: 10.1046/j.1432-1327.1999.00197.x. View

17.

Barbier O, Girard C, Breton R, Belanger A, Hum D . N-glycosylation and residue 96 are involved in the functional properties of UDP-glucuronosyltransferase enzymes. Biochemistry. 2000; 39(38):11540-52. DOI: 10.1021/bi000779p. View

18.

Li C, Wu Q . Adaptive evolution of multiple-variable exons and structural diversity of drug-metabolizing enzymes. BMC Evol Biol. 2007; 7:69. PMC: 1885805. DOI: 10.1186/1471-2148-7-69. View

19.

Barbier O, Belanger A . The cynomolgus monkey (Macaca fascicularis) is the best animal model for the study of steroid glucuronidation. J Steroid Biochem Mol Biol. 2003; 85(2-5):235-45. DOI: 10.1016/s0960-0760(03)00235-8. View

20.

Chen J, Cooper D, Chuzhanova N, Ferec C, Patrinos G . Gene conversion: mechanisms, evolution and human disease. Nat Rev Genet. 2007; 8(10):762-75. DOI: 10.1038/nrg2193. View