Leukotriene D4 receptor-mediated phosphoinositol hydrolysis and calcium mobilization in rat basophilic leukemic cells.
A basophilic leukemic cell line from rat (RBL-1) was used to characterize leukotriene D4 (LTD4) receptor-mediated biochemical and pharmacological effects. [3H]LTD4 binding to the plasma membrane enriched preparation was stereo-selective, specific and saturable. Sodium ions and guanine nucleotides specifically regulated [3H]LTD4 binding to the membrane receptors. Leukotriene E4 (LTE4) and high affinity specific antagonists bound to the receptor with a rank-order potency equivalent to that for the LTD4 receptors in guinea pig lung. In the [3]myoinositol labeled RBL-1 cells, LTD4 and LTE4 induced a rapid hydrolysis of [3H]phosphoinositides. The biosynthesis of the [3H]inositol-trisphosphate was rapid and was detectable at 15-sec poststimulation. The biosynthesis of [3H]inositol-monophosphate was stereo-selective and specific and was inhibited specifically by receptor antagonists. In fura-2 loaded RBL-1 cells, LTD4 and LTE4 induced a transient intracellular Ca++ mobilization. Agonist-induced Ca++ mobilization was specific and stereo-selective and was inhibited by specific receptor antagonists. The most (greater than 85%) LTD4-induced immediate response of Ca++ mobilization was from intracellular sources, whereas a small amount (less than 15%) was derived from the extracellular milieu. Both components were stimulated by receptor agonists and inhibited by the receptor antagonists, suggesting that they were regulated by the LTD4 membrane receptors. In addition, the results also suggested that a guanine nucleotide binding protein, insensitive to islet activating protein from Bordetella pertussis (not Gi or Go), was involved in the signal transduction mechanisms for LTD4 receptors in RBL-1 cells. These results suggested that the plasma membrane enriched LTD4 receptor was coupled via an islet activating protein insensitive G protein to a phosphoinositide specific phospholipase C. Agonist binding to the receptor could activate phospholipase C and resulted in phosphoinositide hydrolysis. Diacylglycerol and inositol trisphosphate could function as intracellular messengers that trigger or contribute to calcium mobilization in RBL-1 cells.
Mong
S
S
Department of Molecular Pharmacology, Smith Kline & French Laboratories, King of Prussia, Pennsylvania.
Wu
H L
HL
Wong
A
A
Sarau
H M
HM
Crooke
S T
ST
D000818:Animals; D001491:Basophils / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D006868:Hydrolysis; D007295:Inositol Phosphates / Q000378:metabolism*; D007942:Leukemia, Experimental / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D018077:Receptors, Leukotriene; D011982:Receptors, Prostaglandin / Q000378:metabolism*; D013189:SRS-A / Q000378:metabolism; D013403:Sugar Phosphates / Q000378:metabolism*
D007295:Inositol Phosphates; D018077:Receptors, Leukotriene; D011982:Receptors, Prostaglandin; D013189:SRS-A; D013403:Sugar Phosphates; D011493:Protein Kinase C; D002118:Calcium