Suppression of cytochrome P450 3A4 function by UDP-glucuronosyltransferase 2B7 through a protein-protein interaction: Cooperative roles of the cytosolic carboxyl-terminal domain and the luminal anchoring region

Yuu Miyauchi, Kiyoshi Nagata, Yasushi Yamazoe, Peter I. Mackenzie, Hideyuki Yamada, Yuji Ishii

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

There is a large discrepancy between the interindividual difference in the hepatic expression level of cytochrome P450 3A4 (CYP3A4) and that of drug clearance mediated by this enzyme. However, the reason for this discrepancy remains largely unknown. Because CYP3A4 interactswith UDP-glucuronosyltransferase 2B7 (UGT2B7) to alter its function, the reverse regulation is expected to modulate CYP3A4-catalyzed activity. To address this issue, we investigated whether protein-protein interaction between CYP3A4 and UGT2B7 modulates CYP3A4 function. For this purpose, we coexpressed CYP3A4, NADPH-cytochrome P450 reductase, and UGT2B7 using a baculovirus-insect cell system. The activity of CYP3A4 was significantly suppressed by coexpressing UGT2B7, and this suppressive effect was lost when UGT2B7 was replaced with calnexin (CNX). These results strongly suggest that UGT2B7 negatively regulates CYP3A4 activity through a protein-protein interaction. To identify the UGT2B7 domain associated with CYP3A4 suppression we generated 12 mutants including chimeras with CNX. Mutations introduced into the UGT2B7 carboxylterminal transmembrane helix caused a loss of the suppressive effect on CYP3A4. Thus, this hydrophobic region is necessary for the suppression of CYP3A4 activity. Replacement of the hydrophilic end of UGT2B7 with that of CNX produced a similar suppressive effect as the native enzyme. The data using chimeric protein demonstrated that the internal membrane-anchoring region of UGT2B7 is also needed for the association with CYP3A4. These data suggest that 1) UGT2B7 suppresses CYP3A4 function, and 2) both hydrophobic domains located near the C terminus and within UGT2B7 are needed for interaction with CYP3A4.

Original languageEnglish
Pages (from-to)800-812
Number of pages13
JournalMolecular Pharmacology
Volume88
Issue number4
DOIs
Publication statusPublished - Oct 1 2015

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Cytochrome P-450 CYP3A
Glucuronosyltransferase
Proteins
Calnexin
NADPH-Ferrihemoprotein Reductase
Baculoviridae
Enzymes

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

Suppression of cytochrome P450 3A4 function by UDP-glucuronosyltransferase 2B7 through a protein-protein interaction : Cooperative roles of the cytosolic carboxyl-terminal domain and the luminal anchoring region. / Miyauchi, Yuu; Nagata, Kiyoshi; Yamazoe, Yasushi; Mackenzie, Peter I.; Yamada, Hideyuki; Ishii, Yuji.

In: Molecular Pharmacology, Vol. 88, No. 4, 01.10.2015, p. 800-812.

Research output: Contribution to journalArticle

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