RT Journal Article
SR Electronic
T1 IDENTIFICATION OF DI-(2-ETHYLHEXYL) PHTHALATE-INDUCED CARBOXYLESTERASE 1 IN C57BL/6 MOUSE LIVER MICROSOMES: PURIFICATION, CDNA CLONING, AND BACULOVIRUS-MEDIATED EXPRESSION
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 1170
OP 1177
DO 10.1124/dmd.104.000620
VO 32
IS 10
A1 Tomomi Furihata
A1 Masakiyo Hosokawa
A1 Nao Koyano
A1 Takahiro Nakamura
A1 Tetsuo Satoh
A1 Kan Chiba
YR 2004
UL http://dmd.aspetjournals.org/content/32/10/1170.abstract
AB Several mouse carboxylesterase (CES) isozymes have been identified, but information about their roles in drug metabolism is limited. In this study, we purified and characterized a mouse CES1 isozyme that was induced by di-(2-ethylhexyl) phthalate. Purified mouse CES1 shared some biological characteristics with other CES isozymes, such as molecular weight of a subunit and isoelectronic point. In addition, purified mouse CES1 behaved as a trimer, a specific characteristic of CES1A subfamily isozymes. The purified enzyme possessed temocapril hydrolase activity, and it was found to contribute significantly to temocapril hydrolase activity in mouse liver microsomes. To identify the nucleotide sequences coding mouse CES1, antibody screening of a cDNA library was performed. The deduced amino acid sequence of the obtained cDNA, mCES1, exhibited striking similarity to those of CES1A isozymes. When expressed in Sf9 cells, recombinant mCES1 showed hydrolytic activity toward temocapril, as did purified mouse CES1. Based on these results, together with the findings that recombinant mouse CES1 had the same molecular weight of a subunit, the same isoelectronic point, and the same native protein mass as those of purified mouse CES1, it was concluded that mCES1 encoded mouse CES1. Furthermore, tissue expression profiles of mCES1 were found to be very similar to those of the human CES1 isozyme. This finding, together with our other results, suggests that mCES1 shares many biological properties with the human CES1 isozyme. The present study has provided useful information for study of metabolism and disposition of ester-prodrugs as well as ester-drugs. The American Society for Pharmacology and Experimental Therapeutics