The importance of hydroperoxide activation for the detection and assay of mammalian 5‐lipoxygenase

FEBS Letters

Samuelsson, Bengt¹  Rouzer, Carol A.¹ 

[1] Department of Physiological Chemistry, Karolinska Institutet, S-104 01 Stockholm, Sweden

关键词: Lipoxygenase Leukotriene Hydroperoxyeicosatetraenoic acid Hydroxyeicosatetraenoic acid Arachidonic acid Inflammation;    5-LO;    5-lipoxygenase;    20:4;    arachidonic acid;    HPETE;    hydroperoxyeicosatetraenoic acid;    HETE;    hydroxyeicosatetraenoic acid;    HPLC;    high pressure liquid chromatography;    DTT;    dithiothreitol;    GSH;    glutathione;   

DOI  :  10.1016/0014-5793(86)80831-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Sulfhydryl reagents such as dithiothreitol stabilized human leukocyte 5-lipoxygenase (5-LO) during purification. During enzyme assay, however, these reagents led to irreproducible or unexpectedly low activity. This inconsistency in the assay was eliminated by inclusion of hydroperoxyeicosatetraenoic acids (1–5 μM) during the reaction which effected a 10–20-fold stimulation of 5-LO activity. Structural studies indicated that an intact hydroperoxy function, and a long-chain fatty acyl moiety were required for 5-LO stimulation. These data suggest that human leukocyte 5-LO is activated by hydroperoxy fatty acids, and that this results in a requirement for exogenous hydroperoxide in the presence of sulfhydryl reagents.

【 授权许可】

Unknown