CHEMORECEPTION IN HYDRA-VULGARIS (ATTENUATA) - INITIAL CHARACTERIZATION OF 2 DISTINCT BINDING-SITES FOR L-GLUTAMIC ACID

To elucidate the relationship between L-glutamic acid and the putative chemoreceptor for glutathione, binding of L-[H-3]glutamate to a crude membrane fraction from Hydra vulgaris (attenuata) has been characterized. The binding of L-[H-3]glutamate was rapid, reversible and saturable. A Scatchard analysis of the specific binding revealed values of 10-mu-M for the dissociation constant (K(d)) and 170 pmol/mg for the maximal capacity of binding sites (B(max)). A maximum of 65% of the specific L-[H-3]glutamate binding was inhibited by the chemostimulatory peptide, glutathione. This glutathione-sensitive glutamate binding presumably represents the association of glutamate with a putative chemoreceptor which modulates feeding behavior in hydra. The remaining 35% of the specific L-[H-3]glutamate binding may be due to a second class of glutamate binding sites which is insensitive to glutathione. The identification of glutathione-insensitive glutamate binding is the first indication of a putative glutamate receptor, which may mediate an action independent of the glutathione-induced feeding response. The glutathione-insensitive and glutathione-sensitive sites must have similar affinities for glutamate since these sites were indistinguishable by Scatchard analysis. A preliminary characterization of the glutathione-insensitive site, performed in the presence of saturating levels of glutathione, revealed inhibition of glutathione-insensitive glutamate binding by kainate and quisqualate, but not by N-methyl-D-aspartate. A comparison of glutamate analogue inhibition of total specific L-[H-3]glutamate binding with analogue inhibition of glutathione-insensitive L-[H-3]glutamate binding suggests that kainate and alpha-aminoadipate may be selective ligands for the glutathione-insensitive and glutathione-sensitive glutamate binding sites, respectively.