ISSN:
1471-4159
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Medicine
Notes:
l-[3H]Glutamate binding sites were solubilized from porcine brain synaptic junctions by Triton X-114 in the presence of KC1. The solubilized binding sites bound l-[3H]glutamate reversibly with Kd and Bmax values of 1.48 ± 0.18 μM and 178.2 ± 15.9 pmol/mg of protein, respectively. These binding sites appeared to be integral membrane glycoproteins, with sugar moieties recognized by wheat germ agglutinin. A 49.3-fold purification of these binding sites was achieved by Triton X-114 solubilization, anion-exchange chromatography, and affinity chromatography using wheat germ agglutinin-Sepharose. The apparent molecular mass of the partially purified binding sites was 620 ± 50 kDa. l-[3H]Glutamate bound to the solubilized preparation could be effectively displaced by agonists of non-N-methyl-d-as-partate (NMDA) l-glutamate receptors but not by NMDA or α-amino-4-phosphonobutyrate. The rank order for the competitive ligands in displacing l-[3H]glutamate was: quisqualate 〉 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid 〉 l-glutamate 〉 kainate.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1471-4159.1991.tb06404.x