Chang Y, Amin J, Weiss D S
Department of Physiology and Biophysics, University of South Florida College of Medicine, Tampa 33612-4799.
Mol Pharmacol. 1995 Mar;47(3):595-602.
The transition metal Zn2+ is differentially distributed in the central nervous system, where it is proposed to be a neuromodulator. One of the documented effects of Zn2+ is the antagonism of gamma-aminobutyric acid (GABA)-mediated synaptic inhibition. This antagonism is presumed to result from a direct interaction of Zn2+ with the GABA receptor/ionophore complex, although the characteristics of Zn2+ sensitivity are dependent on the particular GABA subunit combination. In this study, we examined the effects of Zn2+ on homomeric rho 1 GABA-activated channels expressed in Xenopus oocytes. Zn2+ was found to be a mixed antagonist of these recombinant rho 1 GABA receptors. The antagonism was predominantly competitive at low Zn2+ concentrations (< or = 100 microM), whereas at high Zn2+ concentrations (> 100 microM) a noncompetitive antagonism was apparent. Evidence is presented showing that the antagonism was not due to an interaction of GABA and Zn2+ in solution but, rather, resulted from interactions of these two ligands with the GABA-activated channel. A mechanism is proposed for Zn(2+)-mediated antagonism in which GABA and Zn2+ bind to distinct sites on the GABA complex. The apparent mixed antagonism may arise from different Ki values for the binding of Zn2+ to non-agonist-bound or agonist-bound receptors. However, two distinct Zn2+ binding sites, one competitive and one noncompetitive, could also give rise to the dual antagonism.
过渡金属锌离子(Zn2+)在中枢神经系统中呈差异性分布,据推测它在其中作为一种神经调质发挥作用。锌离子已被证实的作用之一是拮抗γ-氨基丁酸(GABA)介导的突触抑制。这种拮抗作用被认为是锌离子与GABA受体/离子载体复合物直接相互作用的结果,尽管锌离子敏感性的特征取决于特定的GABA亚基组合。在本研究中,我们检测了锌离子对非洲爪蟾卵母细胞中表达的同聚体rho 1 GABA激活通道的影响。结果发现锌离子是这些重组rho 1 GABA受体的混合拮抗剂。在低锌离子浓度(≤100微摩尔)时,拮抗作用主要为竞争性的,而在高锌离子浓度(>100微摩尔)时,非竞争性拮抗作用明显。有证据表明,这种拮抗作用并非由于溶液中GABA与锌离子的相互作用,而是这两种配体与GABA激活通道相互作用的结果。我们提出了一种锌离子介导的拮抗作用机制,其中GABA和锌离子结合到GABA复合物上不同的位点。明显的混合拮抗作用可能源于锌离子与非激动剂结合或激动剂结合受体结合时不同的解离常数(Ki)值。然而,两个不同的锌离子结合位点,一个竞争性的和一个非竞争性的,也可能导致双重拮抗作用。