Role of 5-lipoxygenase-activating protein in the regulation of 5-lipoxygenase activity in human neutrophils

Autor: Hatzelmann Armin, R. Kupferschmidt, K. H. Mohrs, R. Fruchtmann, Reiner Muller-Peddinghaus, R. Kast, S. Raddatz
Rok vydání: 1994
Předmět:
Zdroj: Agents and Actions. 41:C166-C168
ISSN: 1420-908X
0065-4299
DOI: 10.1007/bf01987624
Popis: Using intact and fractionated human polymorphonuclear leukocytes (PMNL), we provide evidence that the enantioselective leukotriene synthesis inhibitor (LSI) BAY X 1005, (R)-2-[4-(quinolin-2-yl-methoxy)phenyl]-2-cyclopentyl acetic acid binds specifically to a high-affinity binding site, which is most likely identical to FLAP. BAY X 1005 blocks the translocation of 5-lipoxygenase (5-LOX) in PMNL stimulated by the calcium ionophore A23187 or chemotactic stimuli such as PAF, C5a or fMLP as does MK-886. In contrast to the direct 5-LOX inhibitors (LOI) A-64077 and AA-861, the degree of leukotriene synthesis inhibition declines with increasing duration of A23187-induced leukocyte activation in the presence of BAY X 1005 and MK-886. Kinetic studies performed with BAY X 1005 showed that this effect was not accompanied by a significant translocation of 5-LOX from the cytosol to the microsomal fraction. Because FLAP has been implicated in the transfer of arachidonic acid to 5-LOX and A23187 is a potent activator of leukocyte phospholipase A2, we hypothesized that the observed loss of leukotriene synthesis inhibition may be due to competition of BAY X 1005 binding by endogenously released arachidonic acid. Accordingly, binding of BAY X 1005 to FLAP in intact and fractionated cells is dose-dependently inhibited by arachidonic acid and other unsaturated long-chain fatty acids, but not by saturated fatty acids. Therefore, we conclude that BAY X 1005 or MK-886 inhibit leukotriene biosynthesis by binding to FLAP, thereby preventing 5-LOX translocation and substrate transfer to the enzyme.
Databáze: OpenAIRE
Externí odkaz: