INHIBITORS FOR CHARACTERIZING TASTE RECEPTORS

味觉接受的最初事件涉及味觉的相互作用 用受体细胞质膜刺激。之前的研究已经 提供的证据表明，对于氨基酸口味的接受，最初的结合 这一过程发生在氨基酸和与膜相关的 蛋白。为了详细定义接收过程，并制定一个 为寻求纠正受体疾病的进一步研究奠定了基础， 这些执行识别功能的蛋白质必须被表征 而且与世隔绝。要做到这一点，需要结合过程的特定抑制剂 被识别或开发。这项建议使用了多学科 识别化合物的方法(神经生理学和生物化学)1) 对味觉中的结合和转导步骤起抑制作用，2) 产生作为激动剂或拮抗剂的单抗 受体结合部位，以及3)抑制物-受体的特征 使用生化和神经生理学程序与 尊重亲和力、专一性和竞争类型。动物模型 用于这项工作的是鲶鱼，因为与陆地脊椎动物不同，它 拥有高密度的受体，对很低的浓度很敏感 氨基酸的浓度，并已被用于许多行为， 神经生理学和生化研究。因此，它的 与味道相关的特征已经有了很好的记录。神经生理学 研究将确定化学抑制剂所表现出的抑制类型 和抑制性抗体，并允许将它们归类为激动剂 或者是敌手。生化研究将确定这些特征 (竞争性-非竞争性)结合试验中的抑制剂和 允许对结合的抑制剂的亲和力进行定量估计 地点。这两种类型的分析都将用于确定 针对不同类别氨基酸受体位置的抑制剂，以及 检查每个位点的对映体专一性。一旦确定，并 它们的活性特征和评估，激动剂和拮抗剂可以 用于寻求分离受体或调节剂的进一步研究 它们所抑制的实体。

The initial event in taste reception involves the interaction of the taste stimulus with the receptor cell plasma membrane. Previous studies have provided evidence that, for amino acid taste reception, the initial binding process takes place between the amino acid and a membrane-associated protein. In order to define the process of reception in detail and lay a foundation for further studies that can seek to correct receptor disease, these proteins that perform the recognition function must be characterized and isolated. To do this, specific inhibitors to the binding process need to be identified or developed. This proposal uses multi-disciplinary approaches (neurophysiology and biochemistry) to 1) identify compounds that act as inhibitors to the binding and transduction steps in taste, 2) produce monoclonal antibodies that act as agonists or antagonists against the receptor binding sites, and 3) characterize the inhibitor-receptor interactions using both biochemical and neurophysiological procedures with respect to affinity, specificity and type of competition. The animal model to be used for this work is the catfish since, unlike land vertebrates, it possesses a high density of receptors, is sensitive to very low concentrations of amino acids and has been used in a number of behavioral, neurophysiological and biochemical studies. Consequently, its taste-related characteristics are well documented. Neurophysiological studies will define the type of inhibition displayed by chemical inhibitors and inhibitory antibodies and allow their classification as either agonists or antagonists. Biochemical studies will define the characteristics (competitive - non-competitive) of the inhibitors in a binding assay and allow quantitative estimates of affinity of the inhibitors for the binding site. Both types of analyses will be used to determine specificity of the inhibitors against different classes of amino acid receptor sites, and to examine the enantiomeric specificity of each site. Once identified and their activity characterized and assessed, the agonists and antagonists can be used in further studies seeking to isolate the receptors or regulatory entities they inhibit.