title: Protein domains, catalytic activity, and subcellular distribution of mouse NTE-related esterase.
abstract: A mammalian family of lipid hydrolases, designated "patatin-like phospholipase domain containing (PNPLA)" recently has attracted attention. NTE-related esterase (NRE) as a member of PNPLA is an insulin-regulated lysophospholipase with homology to neuropathy target esterase (NTE). Mouse NRE (mNRE) has a predicted amino-terminal transmembrane region (TM), a putative regulatory (R) domain, and a hydrophobic catalytic (C) domain. In the current study, we described the expression of green fluorescent protein (GFP)-tagged constructs of mNRE and mutant proteins lacking the specific protein domains. Esterase assays indicated that neither the TM nor R-domain was essential for mNRE esterase activity, but the TM significantly contributed to its activity. Subcellular distribution showed that mNRE was anchored in ER via its TM domain and that its C-domain was associated with ER. Furthermore, experiments involving proteinase treatment revealed that most of mNRE molecule was exposed on the cytoplasmic face of ER membranes. Collectively, our results for the first time revealed the protein domains, catalytic activity, and subcellular location of mNRE and a simplified model for mNRE was proposed.
authors.affiliation: Key Laboratory of Molecular Biology, College of Bio-information, Chongqing University of Posts and Telecommunications, Nan'an District, 400065 Chongqing, People's Republic of China. changpingan@yahoo.com.cn
authors.forename: Ping-An
authors.initials: PA
authors.lastname: Chang
authors.forename: Zhan-Xiang
authors.initials: ZX
authors.lastname: Wang
authors.forename: Ding-Xin
authors.initials: DX
authors.lastname: Long
authors.forename: Wen-Zhen
authors.initials: WZ
authors.lastname: Qin
authors.forename: Yi-Jun
authors.initials: YJ
authors.lastname: Wu
mesh_terms: D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D002462:Cell Membrane / Q000201:enzymology*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013347:Subcellular Fractions
chemical_list: D002265:Carboxylic Ester Hydrolases; D049452:Green Fluorescent Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; C530968:Pnpla7 protein, mouse