[A kinetic scheme of human neutrophil 5-lipoxygenase activity].
In animal cells arachidonic acid is metabolized via the 5-, 12- and 15-lipoxygenase pathways. The kinetic mechanism of action of plant (soya) and animal (reticulocyte) 15-lipoxygenases is now well established. 5-Lipoxygenase possesses, in all probability, the most complex mechanism of activity regulation. At present several effectors of neutrophil 5-lipoxygenase, both cytosolic and membrane-bound ones, have been identified. The molecular and kinetic mechanisms of action of the enzyme are still open to question. A kinetic scheme of regulation of synthesis of arachidonic acid 5-lipoxygenase metabolites which does not exclude the presence of two binding sites on the enzyme molecule, is proposed. Within the framework of this kinetic scheme the enzyme activator complex may be the active form of the enzyme. There is evidence that the curve for the time dependence of 5-HETE accumulation in neutrophils stimulated by the Ca2+ ionophore A23187 has a maximum, while the corresponding curve for the LTB4 accumulation is a curve with saturation. It was shown that an increase in the concentration of exogenous arachidonate induces the synthesis of 5-HETE, whereas the concentration of LTB4 remains practically unchanged. The results of mathematical analysis of the above kinetic scheme and a comparison of experimental and calculated values suggest that the reaction effector, Ca2+, plays a crucial regulatory role in the observed kinetic dependencies reflecting the formation of two sequential products of 5-lipoxygenase oxidation of arachidonate. In this way Ca2+ strongly influences the first step of the reaction, i.e., 5-HETE formation; its effect on the second reaction step (5-HETE conversion into LTA4) is far less apparent.
Sud'ina
G F
GF
Kobel'kov
G M
GM
Barskiĭ
O A
OA
Varfolomeev
S D
SD
D000465:Algorithms; D001094:Arachidonate 5-Lipoxygenase / Q000097:blood*; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000097:blood; D000001:Calcimycin / Q000494:pharmacology; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000097:blood; D007700:Kinetics; D007975:Leukotriene B4 / Q000097:blood; D008433:Mathematics; D008954:Models, Biological; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*
D001095:Arachidonic Acids; D006893:Hydroxyeicosatetraenoic Acids; D007975:Leukotriene B4; D016718:Arachidonic Acid; D000001:Calcimycin; C022022:5-hydroxy-6,8,11,14-eicosatetraenoic acid; D001094:Arachidonate 5-Lipoxygenase
Kineticheskaia skhema deĭstviia 5-lipoksigenazy neĭtrofilov cheloveka.