Displaceable binding of [3H]l-glutamic acid to non-receptor materials.
Displaceable binding of [3H]l-glutamic acid to non-receptor materials.
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[3H]l-谷氨酸与非受体材料的可置换结合。
DOI：
10.1016/0024-3205(86)90244-4
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发表时间：
1986
期刊：
影响因子：
6.1
通讯作者：
Chiu,TH
中科院分区：
文献类型：
--
作者：
Ito,M;Periyasamy,S;Chiu,TH
在四种缓冲液中研究了L-谷氨酸与微离聚管和玻璃的结合。背景与这些材料的结合可以忽略不计，但通过在Tris-HCl和Tris-柠檬酸缓冲液中的离心力或吸力而增加。用HEPES-KOH或三醋酸酯缓冲液时，这种结合作用明显减弱或消除。L可抑制[~3H]L-谷氨酸与微管的结合，但不抑制谷氨酸和天冬氨酸的D-异构体。DL-2-氨基-7-膦庚酸也不抑制该结合。其他低至中等抑制作用的化合物有：N-甲基-D-天冬氨酸、喹乙醇、L-谷氨酸二乙酯、N-甲基-L-天冬氨酸、红藻氨酸和2-氨基-4-磷酸丁酸酯。变性大鼠脑膜可抑制其结合。在Tris-乙酸酯缓冲液中，反复冻融制备的膜可与蛋白质依赖的[~3H]谷氨酸结合。在谷氨酸结合试验中，建议使用三乙酸乙酯或HEPES-KOH缓冲液。如果使用Tris-HCl或Tris-柠檬酸缓冲液，应进行适当的对照实验，以纠正与微隔离管或玻璃纤维过滤器的结合。
[3H]L-glutamic acid binding to microfuge tubes and glass was investigated in four buffers. Background binding to these materials was negligible, but was increased by centrifugation or suction in Tris-HCl and Tris-citrate buffer. This binding was much less or eliminated when HEPES-KOH, or Tris-acetate buffer was used instead.[3H]L-glutamate binding to microfuge tubes was inhibited by L- but not D-isomers of glutamate and aspartate. DL-2-amino-7-phosphonoheptanoic acid also did not inhibit the binding. Other compounds which showed low to moderate inhibition were: N-methyl-D-aspartate, quisqualate, L-glutamic acid diethyl ester, N-methyl-L-aspartate, kainate, and 2-amino-4-phosphonobutyrate. Binding was inhibited by denatured rat brain membranes. A protein-dependent [3H]glutamate binding was obtained with a repeatedly frozen-thawed membrane preparation when binding was done in Tris-acetate buffer. It is recommended that Tris-acetate or HEPES-KOH buffer should be used in the glutamate binding assay. If Tris-HCl or Tris-citrate buffer is used, appropriate control experiment should be done to correct for binding to microfuge tubes or glass fiber filters.