INHIBITORS FOR CHARACTERIZING TASTE RECEPTORS

味觉接受的初始事件涉及味觉的相互作用 刺激受体细胞质膜。 先前的研究 提供的证据表明，对于氨基酸的味觉接收，最初的结合 这个过程发生在氨基酸和膜相关的蛋白质之间。 蛋白 为了详细定义接收过程， 为进一步研究寻求纠正受体疾病奠定了基础， 这些执行识别功能的蛋白质必须被表征 与世隔绝 要做到这一点，结合过程的特定抑制剂需要 被发现或开发。 该提案采用多学科 方法（神经生理学和生物化学）来1）鉴定化合物， 作为味觉中结合和转导步骤的抑制剂，2） 产生单克隆抗体，作为对抗的激动剂或拮抗剂 受体结合位点，和3）表征所述受体-受体 使用生物化学和神经生理学程序与 关于亲和力、特异性和竞争类型。 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， 拥有高密度的受体，对非常低的 浓度的氨基酸，并已用于许多行为， 神经生理学和生物化学研究。 因此它的 与味道相关的特征是有据可查的。 神经生理 研究将确定化学抑制剂显示的抑制类型 和抑制性抗体，并允许将它们分类为激动剂， 或拮抗剂。 生物化学研究将确定 （竞争性-非竞争性）的抑制剂，以及 允许定量估计抑制剂对结合的亲和力 绝佳的价钱 这两种类型的分析将用于确定 针对不同种类的氨基酸受体位点的抑制剂，以及 检查每个位点的对映体特异性。 一旦被发现， 它们的活性被表征和评估，激动剂和拮抗剂可以 用于进一步的研究，寻求分离受体或调节 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.