L-( sup 3 H) glutamate binding to a membrane preparation from the optic lobe of the giant freshwater prawn Macrobrachium rosenbergii de Man
- Mahidol Univ., Bangkok (Thailand) Mahidol Univ., Nakorn Pathom (Thailand)
Membrane preparation from the optic lobe of the giant freshwater prawn, Macrobrachium rosenbergii de Man, was examined for the presence of specific L-({sup 3}H) glutamate binding. The optic lobes were isolated from live animals. The tissue was homogenized and the membrane fraction isolated by differential centrifugation. The membrane suspension was incubated with 10-1,000 nM of L-({sup 3}H) glutamate at 37{degree}C for 60 min. Nonspecific binding was determined by incubating the mixture with 100 {mu}M L-glutamate. L-({sup 3}H) glutamate specifically bound to the membrane fraction with a dissociation equilibrium constant (Kd) of 205 nM and maximum number of binding sites (Bmax) of 2.04 n mol/mg protein. By using LIGAND computerized program, the saturation isotherm binding pattern indicates a single type of binding. To determine the type of glutamate receptors, competitive inhibition and IC{sub 50} of several glutamate agonists and antagonists were determined. The study reveals a metabotropic type of binding site.
- OSTI ID:
- 5857879
- Journal Information:
- Life Sciences; (USA), Vol. 48:1; ISSN 0024-3205
- Country of Publication:
- United States
- Language:
- English
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550201* - Biochemistry- Tracer Techniques