Structural requirements for the binding of fatty acids to 5-lipoxygenase-activating protein.
5-Lipoxygenase-activating protein is required for cellular leukotriene synthesis and is the target of the leukotriene biosynthesis inhibitors MK-886 (3-[1-(p-chlorophenyl)-5-isopropyl-3-tert-butylthio-1H- indol-2-yl]-2,2-dimethylpropanoic acid) and MK-591 (3-[1-(4-chlorobenzyl)-3-(t-butylthio)-5-(quinolin-2-ylmethoxy)-indol-2-yl] - 2,2-dimethylpropanoic acid). Recent studies demonstrate that 5-lipoxygenase-activating protein binds arachidonic acid and stimulates the utilization of this substrate by 5-lipoxygenase. The present study utilizes a radioligand binding assay to assess the affinity of 5-lipoxygenase-activating protein for arachidonic acid and the specificity of the fatty acid binding site on 5-lipoxygenase-activating protein. Our findings demonstrate that the presence of a free carboxyl group on fatty acids or leukotriene biosynthesis inhibitors which interact with 5-lipoxygenase-activating protein is not required for specific binding to the protein. However, the degree of saturation significantly affects the affinity of fatty acids for 5-lipoxygenase-activating protein.
Charleson
S
S
Department of Biochemistry, Merck Frosst Centre For Therapeutic Research, Pointe Claire-Dorval, Quebec, Canada.
Evans
J F
JF
Léger
S
S
Perrier
H
H
Prasit
P
P
Wang
Z
Z
Vickers
P J
PJ
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D058927:5-Lipoxygenase-Activating Proteins; D016718:Arachidonic Acid / Q000378:metabolism*; D001665:Binding Sites / Q000187:drug effects; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology; D007211:Indoles / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism*; D015289:Leukotrienes / Q000096:biosynthesis; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D011804:Quinolines / Q000378:metabolism; D011869:Radioligand Assay; D002722:Chlorobenzenes
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D058927:5-Lipoxygenase-Activating Proteins; D016718:Arachidonic Acid; D002352:Carrier Proteins; D006893:Hydroxyeicosatetraenoic Acids; D007211:Indoles; D015289:Leukotrienes; D008565:Membrane Proteins; D011804:Quinolines; C548592:ALOX5AP protein, human; C060893:MK-886; D002722:Chlorobenzenes; C078266:MK 0591; C074616:L 691831; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid