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Murine 12(R)-lipoxygenase: functional expression, genomic structure and chromosomal localization
1Research Program on Tumor Cell Regulation, Deutsches Krebsforschungszentrum, Heidelberg, Germany. p.krieg@dkfz-heidelberg.de
FEBS Letters
|April 1, 1999
Summary
Researchers identified a mouse skin cDNA encoding 12(R)-lipoxygenase. This enzyme specifically converts methyl arachidonate to 12-HETE, with its gene located on mouse chromosome 11.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Lipoxygenases are enzymes involved in fatty acid metabolism.
- A novel lipoxygenase from mouse skin was recently cloned.
Purpose of the Study:
- To characterize the enzymatic activity and gene structure of the cloned 12(R)-lipoxygenase.
- To determine the substrate specificity and stereochemical outcome of the enzymatic reaction.
- To map the gene's chromosomal location.
Main Methods:
- cDNA cloning and expression in HEK cells.
- Enzymatic assays using methyl arachidonate, arachidonic acid, and linoleic acid.
- Chiral phase chromatography for enantiomer determination.
- Gene structure analysis (exons/introns).
- Interspecific backcross analysis for gene mapping.
Main Results:
- The cloned cDNA encodes a functional 12(R)-lipoxygenase.
- The recombinant enzyme specifically converts methyl arachidonate to the R-enantiomer of 12-HETE.
- Arachidonic acid and linoleic acid are not substrates.
- The 12(R)-lipoxygenase gene has a unique structure with 15 exons and 14 introns.
- The gene is localized to mouse chromosome 11.
Conclusions:
- The cloned mouse cDNA encodes a novel 12(R)-lipoxygenase with specific substrate preference.
- The unique gene structure and chromosomal localization provide insights into lipoxygenase evolution and function.
- This enzyme represents a distinct pathway in eicosanoid metabolism.