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1.
FEBS Lett ; 445(2-3): 301-5, 1999 Feb 26.
Article in English | MEDLINE | ID: mdl-10094477

ABSTRACT

Endogenous, constitutive soluble epoxide hydrolase in mice 3T3 cells was localized via immunofluorescence microscopy exclusively in peroxisomes, whereas transiently expressed mouse soluble epoxide hydrolase (from clofibrate-treated liver) accumulated only in the cytosol of 3T3 and HeLa cells. When the C-terminal lie of mouse soluble epoxide hydrolase was mutated to generate a prototypic putative type 1 PTS (-SKI to -SKL), the enzyme targeted to peroxisomes. The possibility that soluble epoxide hydrolase-SKI was sorted slowly to peroxiosmes from the cytosol was examined by stably expressing rat soluble epoxide hydrolase-SKI appended to the green fluorescent protein. Green fluorescent protein soluble epoxide hydrolase-SKI was strictly cytosolic, indicating that -SKI was not a temporally inefficient putative type 1 PTS. Import of soluble epoxide hydrolase-SKI into peroxisomes in plant cells revealed that the context of -SKI on soluble epoxide hydrolase was targeting permissible. These results show that the C-terminal -SKI is a non-functional putative type 1 PTS on soluble epoxide hydrolase and suggest the existence of distinct cytosolic and peroxisomal targeting variants of soluble epoxide hydrolase in mouse and rat.


Subject(s)
Epoxide Hydrolases/metabolism , 3T3 Cells , Animals , Epoxide Hydrolases/genetics , HeLa Cells , Humans , Isoenzymes/genetics , Isoenzymes/metabolism , Mammals , Mice , Microbodies/enzymology , Rats , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Solubility , Subcellular Fractions , Transfection
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