Purification and molecular characterization of the brain synaptic membrane glutamate-binding protein.
A glutamate-binding protein from rat brain synaptic plasma membranes has been purified to apparent homogeneity. This protein has a Mr of 14,300 based on amino acid and carbohydrate analyses. The protein is enriched with tryptophan residues, which contribute substantially to its hydrophobic nature. It also has a relatively high content of acidic amino acids, which determine is low isoelectric point (4.82). The protein exhibits either a single, high-affinity class of sites for L-[3H]glutamate binding (KD = 0.13 microM) when binding is measured at low protein concentrations, or two classes of sites with high (KD = 0.17 microM) and low affinities (KD = 0.8 microM) when binding is measured at high protein concentrations. These observations suggest preferential binding of L-glutamate to a self-associating form of the protein. The displacement of protein-bound L-[3H]glutamic acid by other neuroactive amino acids has characteristics similar to those observed for displacement of L-glutamate from membrane binding sites. Chemical modification of the cysteine and arginine residues results in an inhibition of glutamate binding activity. The possible function of this protein in the physiologic glutamate receptor complex of neuronal membranes is discussed.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002241:Carbohydrates / Q000032:analysis; D002352:Carrier Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D007525:Isoelectric Focusing; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D017470:Receptors, Glutamate; D013570:Synaptic Membranes / Q000378:metabolism*; D013696:Temperature
D000596:Amino Acids; D002241:Carbohydrates; D002352:Carrier Proteins; D005971:Glutamates; D011956:Receptors, Cell Surface; D017470:Receptors, Glutamate; D018698:Glutamic Acid