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 isoform 、 Intron 、 Alternative splicing 、 RNA splicing 、 Chemistry 、 Biochemistry 、 Arachidonate 5-lipoxygenase 、 Exon 、 Enzyme activator 、 Isozyme
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.