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Winkelhake JL, Voss W Jr, Lopatin DE.
“Comparative Inhibitory Action of D- and L-Tryptophan on the Effect of d-Lysergic Acid Diethylamide in Vitro.”.
Molec Pharmacol.. 1974;10(1):68-77.
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Abstract
The inhibitory effect of D- and L-tryptophan on incorporation in vitro of d-(3H) LSD into protein of hyperimmune rabbit lymphoid cells, was studied. Methods Adult albino rabbits were immunised intrascapularly and in the hind footpads with a total of 10 mg emulsified fluorescyl-conjugated porcine y-globulin. Blood samples were obtained and rabbits killed 4-6 days later. Lymphoid cells were extracted from spleen and or popliteal lymph nodes- and incubated with (3H) leucine, (4C)-D-tryptophan ('4C) - L-tryptophan and (3H)-LSD (10-20 mcl). Antibody producing cells were pelleted and dissolved in tissue stabiliser. Samples were counted in a liquid scintillation counter. When LSD was incubated with immune rabbit lymphoid cells in the presence of 3H-leucine and 4C tryptophan, tryptophan incorporation was inhibited. When D-tryptophanwas added to cells incubated with LSD, interference was abolished. L-tryptophan did not have this effect. As radioactivity from labeled LSD was released on incubation with Pronase, it was concluded that LSD was incorporated covalently into protein. 3H-LSD incorporation was inhibited by puromycin, suggesting that this incorporation was an active process. Tyrosine, proline and histidine did not inhibit incorporation but lysergic acid, indoleacetic acid (IAA), DL-7-azatryptophan and tryptazan did. Anti-LSD antibodies bound secreted peptides as did anti-IAA antibodies Material secreted from antibody producing cells, when incubated with excess D-or L-tryptophan corresponded to the mol. wt of immunoglobulin. Antibody producing cells were incubated in minimal essential medium minus tryptophan and in 20% and 50% rabbit serum. All radioactivity found in the extracellular fraction was inhibited by puromycin and D-tryptophan, thus levels of free tryptophan did not inhibit LSD effect in vitro at drug concentrations of 2-4mcg/ml. There were no significant differences in 3H-accumulation after incubation of immune cells with and without D or L-tryptophan or L-histidine. From these results it is possible to formulate the preliminary steps culminating ín the covalent incorporation of LSD into synthesised protein.
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