Molecular cloning and expression of the C-terminal domain of mouse NTE-related esterase.
NTE-related esterase (NRE), conserved in mouse, rat and human, was a member of patatin-like phospholipases (PLPLA) with high homology to neuropathy target esterase (NTE). Little has been known about the characteristics of NRE and NRE functional esterase activity has yet not been defined. The C-terminal gene sequence of mouse NRE (mNREC) encoding 923-1,326 amino acid containing the patatin domain was first cloned and then expressed tagged with enhanced green fluorescence protein (EGFP) in mammalian cells. The results showed that mNREC had NTE esterase activity in mammalian cells. Overexpression of mNREC did not affect the esterase activity sensitive to paraoxon or resistant to both paraoxon and mipafox. mNREC was distributed in the cytoplasm in contrast to the distribution of human NTE esterase domain. The expression analysis of NRE gene in adult mouse tissues by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) showed that there were higher levels of NRE mRNA in the brain and testis than in the liver and kidney, which was about 50% and 35% of that in the brain. These results firstly showed the tissue distribution of NRE gene in adult mouse and defined that NRE had functional esterase activity.
Chang
Ping-An
PA
Key Laboratory of Molecular Biology, College of Bio-information, Chongqing University of Posts and Telecommunications, Chongqing 400065, PR China. changpingan@yahoo.com.cn
Long
Ding-Xin
DX
Wu
Yi-Jun
YJ
D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002265:Carboxylic Ester Hydrolases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic; D007531:Isoflurophate / Q000031:analogs & derivatives / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010261:Paraoxon / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D014631:Valerates / Q000378:metabolism
D008565:Membrane Proteins; D012333:RNA, Messenger; D014631:Valerates; D007531:Isoflurophate; C049867:phenyl valerate; C005238:mipafox; D002265:Carboxylic Ester Hydrolases; D010261:Paraoxon