Novel 5-lipoxygenase isoforms affect the biosynthesis of 5-lipoxygenase products

作者: Luc H. Boudreau , Jonathan Bertin , Philippe P. Robichaud , Mark Laflamme , Rodney J. Ouellette

DOI: 10.1096/FJ.10-173856

关键词: Gene isoformIntronAlternative splicingRNA splicingChemistryBiochemistryArachidonate 5-lipoxygenaseExonEnzyme activatorIsozyme

摘要: 5-Lipoxygenase (5-LO) is the essential enzyme for biosynthesis of leukotrienes, important mediators inflammation. This study investigated whether variants 5-LO exist in human leukocytes. mRNA isoforms that are consistent with alternative splicing were identified by RT-PCR a cell line or type-specific pattern. All evaluated cells expressed containing all 14 exons expected sites. Individual retained intron 10 (α-10), lacked exon 13 (Δ-13), and (Δ-10,13) first 96 base pairs (Δ-p10) identified. Immunoreactive bands coeluting cloned α-10 Δ-13 measured primary neutrophils Raji cells. When HEK293 cells, proteins without catalytic activity. However, when coexpressed active full-length 5-LO, significantly decreased products up to 44%, as assessed reverse-phase HPLC analysis. Additionally, stimulated was detected immunoblot both nuclear non-nuclear compartments, while isoform only fraction. These may represent new mechanism regulation pathway lipid mediator biosynthesis.

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