Receptor Proteins in Vertebrate Olfaction

脊椎动物嗅觉中的受体蛋白

• 批准号: 8820690
• 负责人: Steven Price
• 金额: $ 10.8万
• 依托单位: Virginia Commonwealth University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-01 至 1992-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8820690&HistoricalAwards=false
• 关键词: Receptor; Proteins; Vertebrate; Olfaction

The long term objective of Dr. Price's research is to understand the mechanism (or mechanisms) by which chemicals in the air stimulate olfactory receptor cells in the nose, thus leading to odor perception. One mechanism suggested is that the odorous chemicals (odorants) bind to specific receptor proteins on the surface of the cells, which causes the cells to generate nerve impulses. This research project will test the hypothesis that certain proteins isolated from the tissue containing olfactory receptor cells are, in fact, receptor proteins and a determination of the protein characteristics will be made. Isolating the proteins by affinity chromatography takes advantage of the fact that they bind certain odorants. Odorant analogues are linked to solid supports and made into columns through which tissue extracts are passed. Proteins that bind odorants often bind the analogues as well. The analogues are linked to the solid support, thus these proteins are separated from the thousands of other chemicals in the extracts. The proteins are recovered by washing the solid support-analogue-protein complex with concentrated solutions of the analogue. Binding an odorant does not prove that a protein is a receptor. This problem is approached by preparing antibodies that interact with different parts of the protein (so-called monoclonal antibodies). The antibodies are applied to olfactory receptor tissue and responses to odorants are measured. If the protein is a receptor, some of the antibodies should prevent it from binding the odorant and, therefore, inhibit responses. Two proteins have been isolated from olfactory tissue and subjected to this test. The results suggest that they are receptors. Receptor proteins are hypothesized to comprise a family of similar proteins. Some antibodies against each of the two proteins mentioned inhibit responses to several unrelated odorants. This finding is consistent with the stated hypothesis; the antibodies probably interact with part of the protein that is common to all receptor proteins. The two proteins are indistinguishable by electrophoresis, which estimates protein size on the basis of the rate at which it moves in an electric field. This, too, is consistent with the hypothesis that receptor proteins comprise a structural class. These studies are intended to elaborate upon the investigations described above by seeking and comparing additional proteins. Samples will be provided to other investigators for use in their experiments on olfactory mechanisms.

普莱斯博士研究的长期目标是了解 空气中的化学物质 刺激鼻子中的嗅觉受体细胞，从而导致 气味感知 其中一种机制是， 化学物质（气味剂）结合到特定的受体蛋白上， 细胞的表面，这使得细胞产生神经 冲动 这个研究项目将检验这样一个假设， 某些蛋白质是从含有嗅觉的组织中分离出来的， 事实上，受体细胞是受体蛋白， 将进行蛋白质特性的测定。 通过亲和层析分离蛋白质利用了 它们能结合某些气味。 气味类似物 连接到固体支撑物上并制成柱子， 组织提取物通过。 结合气味物质的蛋白质通常 也会结合类似物。 类似物与 固体支持物，因此这些蛋白质从 提取物中含有数千种其他化学物质。 将蛋白质 通过洗涤固体载体-类似物-蛋白质复合物回收 用类似物的浓缩溶液。 与气味物质结合并不能证明蛋白质就是受体。 解决这个问题的方法是制备抗体， 蛋白质的不同部分（所谓的单克隆 抗体）。 将抗体应用于嗅觉受体 测量组织和对气味剂的反应。 如果蛋白质是 一些抗体会阻止它与受体结合 从而抑制反应。 两种蛋白质具有 从嗅觉组织中分离出来并进行这项测试。 结果表明，它们是受体。 受体蛋白质被假设为包括一个家族， 相似的蛋白质 一些抗体分别针对这两种 所提到的蛋白质抑制对几种无关的 气味剂。 这一发现与所述假设一致; 抗体可能会与蛋白质的一部分相互作用 所有受体蛋白都是如此。 这两种蛋白质通过电泳无法区分， 根据蛋白质移动的速度来估计蛋白质的大小 在电场中。 这也是符合 假设受体蛋白包含一个结构类别。 这些研究的目的是详细说明调查 通过寻找和比较另外的蛋白质来进行上述描述。 样本将提供给其他研究者，供其 嗅觉机制的实验。