Chen Z, Lancet D
Proc Natl Acad Sci U S A. 1984 Mar;81(6):1859-63. doi: 10.1073/pnas.81.6.1859.
In search for olfactory receptor molecules, we carried out comprehensive electrophoretic mapping of membrane proteins in the cilia of frog olfactory epithelium. Seven polypeptides, extracted from isolated cilia by nonionic detergent, were unique to the sensory organelles, compared to nonsensory (respiratory) counterparts. Olfactory cilia contained 3-10 times more membrane-associated protein as compared to respiratory cilia, in agreement with reported densities of freeze-fracture intramembranous particles. Four of the olfactory polypeptides were major constituents of the ciliary membrane, each amounting to greater than 10% of its total protein. Three major and one minor specific polypeptide were glycosylated, whereas membranes of nonsensory cilia were practically devoid of glycoproteins. A clear difference in surface composition was also shown by microscopic visualization of fluoresceinated lectin bound to intact isolated cilia. Two of the olfactory glycoproteins displayed pronounced heterogeneity of apparent molecular weight, which could partly be due to protein sequence diversity, as expected for odorant receptor molecules. The recently described inhibition of odorant-evoked sensory potentials by the lectin concanavalin A is consistent with the hypothesis that one or more of the specific glycoproteins described here plays a role in olfactory reception.
为了寻找嗅觉受体分子,我们对青蛙嗅觉上皮纤毛中的膜蛋白进行了全面的电泳图谱分析。与非感觉(呼吸)对应物相比,从分离的纤毛中用非离子去污剂提取的七种多肽是感觉细胞器所特有的。嗅觉纤毛所含的膜相关蛋白比呼吸纤毛多3至10倍,这与报道的冷冻断裂膜内颗粒密度一致。其中四种嗅觉多肽是纤毛膜的主要成分,每种多肽占其总蛋白的比例均超过10%。三种主要的和一种次要的特异性多肽被糖基化,而非感觉纤毛的膜实际上不含糖蛋白。通过与完整分离的纤毛结合的荧光素化凝集素的显微镜观察,也显示出表面组成的明显差异。其中两种嗅觉糖蛋白表现出明显的表观分子量异质性,这部分可能是由于蛋白质序列多样性,正如气味受体分子所预期的那样。最近描述的伴刀豆球蛋白A对气味诱发的感觉电位的抑制作用与以下假设一致,即这里描述的一种或多种特异性糖蛋白在嗅觉接收中起作用。