Partial characterization and detergent solubilization of the putative glutathione chemoreceptor from hydra.
Feeding behavior in hydra is initiated by the association of glutathione (GSH) with a putative external chemoreceptor. In the present study, the binding of [35S]GSH to hydra membranes has been characterized. Nondisplaceable [35S]GSH binding which compromised previous analyses [Grosvenor, W., Bellis, S., Kass-Simon, G., & Rhoads, D. (1992) Biochim. Biophys. Acta (in press)] was eliminated by treating membranes with an inhibitor of GSH metabolism, borate in combination with L-serine. The specific binding which was not inhibited by borate/serine demonstrated many of the characteristics expected of a ligand/receptor interaction. The binding was rapid, reversible, and saturable. A Scatchard analysis of saturation isotherms indicated a dissociation constant (KD) of 3.4 microM, a value which is in good agreement with concentrations of glutathione which are known to induce feeding behavior. Hydra membranes were detergent-solubilized with 10 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 100 mM KCl, and 10% glycerol. The soluble fraction contained 40% of the original saturable, reversible GSH binding activity. The KD for GSH binding to the solubilized preparation was estimated as 2.7 microM, a valuable which is not appreciably different from the KD for binding to intact membranes. The fidelity of GSH binding in the solubilized preparation suggests that this preparation will be useful in further characterization of the putative glutathione chemoreceptor.
Department of Biochemistry and Biophysics, University of Rhode Island, Kingston 02881.
S L
SL
Bellis
G
G
Kass-Simon
D E
DE
Rhoads
D000818:Animals; D001888:Boric Acids / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002628:Chemoreceptor Cells / Q000737:chemistry / Q000378:metabolism*; D002793:Cholic Acids; D003902:Detergents; D005247:Feeding Behavior / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism*; D006829:Hydra; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D018000:Receptors, Peptide; D012694:Serine / Q000494:pharmacology; D012995:Solubility; D005723:gamma-Glutamyltransferase / Q000037:antagonists & inhibitors
D001888:Boric Acids; D002793:Cholic Acids; D003902:Detergents; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C035560:glutathione receptor; D012694:Serine; D005723:gamma-Glutamyltransferase; D005978:Glutathione; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; C032688:boric acid