title: 5-lipoxygenase-activating protein is an arachidonate binding protein.
abstract: 5-Lipoxygenase-activating protein (FLAP) is an 18-kDa integral membrane protein which is essential for cellular leukotriene (LT) synthesis, and is the target of LT biosynthesis inhibitors. However, the mechanism by which FLAP activates 5-LO has not been determined. We have expressed high levels of human FLAP in Spodoptera frugiperda (Sf9) insect cells infected with recombinant baculovirus, and used this system to demonstrate that FLAP specifically binds [125I]L-739,059, a novel photoaffinity analog of arachidonic acid. This binding is inhibited by both arachidonic acid and MK-886, an LT biosynthesis inhibitor which specifically interacts with FLAP. These studies suggest that FLAP may activate 5-LO by specifically binding arachidonic acid and transferring this substrate to the enzyme.
authors.affiliation: Department of Pharmacology, Merck Frosst Centre for Therapeutic Research, Quebec, Canada.
authors.forename: J A
authors.initials: JA
authors.lastname: Mancini
authors.forename: M
authors.initials: M
authors.lastname: Abramovitz
authors.forename: M E
authors.initials: ME
authors.lastname: Cox
authors.forename: E
authors.initials: E
authors.lastname: Wong
authors.forename: S
authors.initials: S
authors.lastname: Charleson
authors.forename: H
authors.initials: H
authors.lastname: Perrier
authors.forename: Z
authors.initials: Z
authors.lastname: Wang
authors.forename: P
authors.initials: P
authors.lastname: Prasit
authors.forename: P J
authors.initials: PJ
authors.lastname: Vickers
mesh_terms: D058927:5-Lipoxygenase-Activating Proteins; D000345:Affinity Labels; D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D001095:Arachidonic Acids / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004789:Enzyme Activation; D015870:Gene Expression; D006801:Humans; D007211:Indoles / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009036:Moths / Q000378:metabolism; D010777:Photochemistry; D011804:Quinolines / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
chemical_list: D058927:5-Lipoxygenase-Activating Proteins; C548592:ALOX5AP protein, human; D000345:Affinity Labels; D001095:Arachidonic Acids; D002352:Carrier Proteins; D007211:Indoles; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D011804:Quinolines; D011994:Recombinant Proteins; C074616:L 691831; C079922:L 739059; D001094:Arachidonate 5-Lipoxygenase