Evidence that barbiturates inhibit antigen-induced responses through interactions with a GTP-binding protein in rat basophilic leukemia (RBL-2H3) cells.
Little is known about the mechanism of action of anesthetics at the biochemical level. The present work, however, gives evidence that barbiturates inhibit inositol phospholipid hydrolysis in both intact and permeabilized rat basophilic leukemia (RBL-2H3) cells by an effect on GTP-binding proteins (G-proteins). Inhibition of hydrolysis was observed when intact cells were stimulated with antigen (DNP24 BSA) or with oligomers of IgE. The inhibition was dependent on the concentration and type of barbiturate used with an order of inhibitory action of secobarbital less than S(-) pentobarbital less than pentobarbital less than R(+) pentobarbital less than phenobarbital. The relatively inactive analogue, (1'RS, 3'SR) 3-hydroxypentobarbital caused little (less than 30% at 1 mM) or no inhibition (at 0.1-0.5 mM). In permeabilized cells, the hydrolysis induced by DNP24 BSA and the nonhydrolyzable analogue of GTP, GTP gamma S (2-100 microM), was also inhibited by pentobarbital. The inhibition of hydrolysis was decreased as pH increased, and was no longer apparent at pH 7.8, a possible indication that the inhibitory effect was due to the unionized form of the drug. In permeabilized cells, the inhibition by pentobarbital occurred in the presence or absence of Ca2+ and was uncompetitive in nature (Km = 7.1 microM for GTP in controls vs. 1.6 microM in the presence of 0.5 mM pentobarbital). Taken together, the data suggest that barbiturates alter the activity of G-proteins independently of Ca2+, and the inhibition may depend on both the hydrophobic properties and the stereospecific and structural features of the molecule.
Robinson-White
A J
AJ
Department of Anesthesiology, Uniformed Services University of the Health Sciences, National Institutes of Health, Bethesda, Maryland.
Muldoon
S M
SM
Elson
L
L
Collado-Escobar
D M
DM
D000818:Animals; D000941:Antigens / Q000276:immunology*; D001463:Barbiturates / Q000494:pharmacology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D004140:Dinitrophenols / Q000276:immunology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D006868:Hydrolysis; D007073:Immunoglobulin E / Q000494:pharmacology; D007295:Inositol Phosphates / Q000378:metabolism; D015471:Leukemia, Basophilic, Acute / Q000378:metabolism*; D010424:Pentobarbital / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000378:metabolism; D051381:Rats; D012631:Secobarbital / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000276:immunology; D013873:Thionucleotides / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000941:Antigens; D001463:Barbiturates; D004140:Dinitrophenols; D007295:Inositol Phosphates; D010716:Phosphatidylinositols; D013873:Thionucleotides; C034638:dinitrophenyl-bovine serum albumin; D012631:Secobarbital; D012710:Serum Albumin, Bovine; D007073:Immunoglobulin E; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D010424:Pentobarbital