The importance of hydroperoxide activation for the detection and assay of mammalian 5‐lipoxygenase
- 18 August 1986
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 204 (2) , 293-296
- https://doi.org/10.1016/0014-5793(86)80831-6
Abstract
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.Keywords
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