Spermine modulation of specific [H-3]-gabapentin binding to the detergent-solubilized porcine cerebral cortex alpha(2)delta calcium channel subunit
Spermine modulation of specific [H-3]-gabapentin binding to the detergent-solubilized porcine cerebral cortex alpha(2)delta calcium channel subunit
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DOI：
10.1038/sj.bjp.0700988
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发表时间：
1997-03-01
影响因子：
7.3
通讯作者：
Woodruff, GN
中科院分区：
文献类型：
--
作者：
Dissanayake, VUK;Gee, NS;Woodruff, GN
1最近的研究已经鉴定了从猪大脑皮层膜纯化的[H-3]-加巴喷丁结合蛋白作为电压敏感性钙通道的α（2）δ亚基（Gee等人，1996年）。本研究探讨了多胺精胺对特异性[H-3]-加巴喷丁与洗涤剂溶解的猪大脑皮层膜结合的影响。2精胺、亚精胺、1，10-癸二胺、Mg 2+和Zn 2+（所有二价阳离子）以浓度依赖性方式取代[H-3]-加巴喷丁与洗涤剂溶解的膜结合，最大抑制率为65- 75%。放射性配体结合研究表明，精胺不直接与[H-3]-加巴喷丁结合位点相互作用。精胺抑制[H-3]-加巴喷丁的结合，通过与膜蛋白上的多胺敏感的变构位点相互作用。精胺抑制[H-3]-加巴喷丁结合的陡峭浓度依赖性可能表明多位点协同性。3大脑皮质膜和吐温20增溶膜的延长透析导致[H-3]-加巴喷丁结合增加>2.0倍。结合的增加是由于去除了热稳定的低分子量（
1 Recent studies have identified the [H-3]-gabapentin-binding protein, purified from porcine cerebral cortical membranes, as the alpha(2) delta subunit of voltage-sensitive calcium channels (Gee et al., 1996). The present study investigates the influence of the polyamine spermine on specific [H-3]-gabapentin binding to detergent-solubilized porcine cerebral cortical membranes.2 Spermine, spermidine, 1,10 diaminodecane, Mg2+ and Zn2+, all divalent cations, displaced [H-3]gabapentin binding to detergent-solubilized membranes in a concentration-dependent manner with a maximal inhibition of 65-75%. Radioligand binding studies showed that spermine did not directly interact with the [H-3]-gabapentin-binding site. Spermine inhibited [H-3]-gabapentin binding by interacting with a polyamine-sensitive allosteric site on the membrane protein. The steep concentration-dependence of spermine inhibition of [H-3]-gabapentin binding may suggest multi-site co-operativity.3 Prolonged dialysis of cerebral cortical membranes and Tween 20-solubilized membranes resulted in a >2.0 fold increase in [H-3]-gabapentin binding. The increase in binding was due to the removal of a heat stable, low molecular weight (