Identification and characterisation of soluble epoxide hydrolase in mouse brain by a robust protein biochemical method.
The central nervous system is an important potential target for certain environmental prototoxins, but relatively little is known regarding brain-specific expression of biotransformation enzyme systems. On the other hand, developments in the field of molecular biology and advances in high-throughput screening methods continue to increase the number and amounts of available proteins. We used thus a robust and reliable technique, two-dimensional gel electrophoresis coupled to matrix assisted laser desorption/ionisation mass spectroscopy followed by tandem mass spectrometry and identified for the first time soluble epoxide hydrolase and added other biotransformation enzymes in the hippocampal region of mouse brain. Soluble epoxide hydrolase has an Mr of 61.5 kDa, pI of 5.9, twenty-six matching peptides and sequence coverage of 56% and was unambiguously identified by MS/MS. Since localised biotransformation events in regions of the central nervous system may account for pathologies and/or toxicities initiated by exposure to certain endogenous and/or environmental chemicals, identification of these enzymes would present an opportunity for developing novel therapeutic targets or would have critical toxicologic significance.
Shin
J-H
JH
Department of Pediatrics, Medical University of Vienna, 1090 Vienna, Austria.
Engidawork
E
E
Delabar
J-M
JM
Lubec
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004798:Enzymes / Q000032:analysis; D004851:Epoxide Hydrolases / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D004798:Enzymes; D004851:Epoxide Hydrolases