Characterization of L-( sup 3 H)glutamate binding sites in the synaptic membranes of rat brain

Thesis/Dissertation · Fri Jan 01 00:00:00 EST 1988

OSTI ID:5413546

Periyasamy, S

In this study we have shown that L-({sup 3}H) glutamate binds to microfuge tubes and glass fiber filters in the presence of Tris-HCl or Tris-citrate buffer with high affinity, stereospecificity and in a saturable manner; and this binding was inhibited by glutamate analogs. A non-linear protein-dependent binding was observed when binding was carried out with four-times washed crude synaptic membranes in microfuge tubes and in Tris-HCl buffer. However, the binding was found to be linear when repeatedly frozen-thawed and washed membranes were used. Further, repeated freezing-thawing and washing produced an increase in L-({sup 3}H)glutamate binding. The supernatants obtained from repeated freezing-thawing inhibited L-({sup 3}H)glutamate binding and repeated freezing-thawing and washing procedure seemed to be very effective in removing endogenous inhibitor(s) of glutamate binding. Scatchard analysis of specific binding showed a single class of binding sites in four-times washed crude synaptic membranes.

• Research Organization: Medical Coll. of Ohio, Toledo, OH (USA)
• OSTI ID: 5413546
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GLUTAMIC ACID
RECEPTORS
CHEMICAL COMPOSITION
BIOCHEMICAL REACTION KINETICS
BRAIN
CELL MEMBRANES
FREEZING
NERVE CELLS
RATS
THAWING
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINO ACIDS
ANIMAL CELLS
ANIMALS
BODY
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
KINETICS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
REACTION KINETICS
RODENTS
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques