An LTC4 binding site in gastric mucosa is shared with glutathione.
Recent results from our laboratory and others have suggested a possible physiological functional role(s) for leukotrienes in gastric mucosa. In the present study 3H-LTC4 binds to washed rabbit gastric mucosal membranes at 4 degrees C with a Kd of 5 nM and a Bmax of 31.3 pmol/mg protein. Leukotrienes D4, E4, B4, oxidized glutathione (GSSG), cysteine, and mercaptoethanol were unable to displace 3H-LTC4 at 1 microM concentrations, while GSH inhibited binding with a Ki of 47 nM. Differential centrifugation of the membrane preparation to remove mitochondria resulted in Ki values for LTC4 and GSH of 14 and 23 nM, respectively. The similar binding affinities and competitive receptor binding kinetics for GSH and LTC4, the low affinity for other leukotrienes, and a Ki of 7 microM for hematin, a substrate for glutathione S-transferase, suggest that 3H-LTC4 binds to a GSH site which does not discriminate between LTC4 and GSH. Membranes fractionated to remove mitochondria were assayed for glutathione peroxidase, gamma-glutamyltranspeptidase, and glutathione S-transferase as possible binding sites for LTC4. We were unable to detect enzyme activity for any of the three enzymes. The binding of LTC4 in gastric mucosa differs from other tissues with respect to the high affinity for GSH, and thus becomes an appropriate tissue in which to investigate the relationships between LTC4 and GSH.
Pendley
C E
CE
Rorer Central Research, Horsham, PA 19044.
Pendleton
R G
RG
Herczeg
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T
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D005260:Female; D005753:Gastric Mucosa / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006427:Hemin / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D011817:Rabbits; D011971:Receptors, Immunologic / Q000378:metabolism*; D018077:Receptors, Leukotriene; D013189:SRS-A / Q000378:metabolism*; D013696:Temperature; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D008565:Membrane Proteins; D011971:Receptors, Immunologic; D018077:Receptors, Leukotriene; D013189:SRS-A; D006427:Hemin; D005979:Glutathione Peroxidase; D005723:gamma-Glutamyltransferase; D005982:Glutathione Transferase; D005978:Glutathione