Goldstein N I, Cagan R H
Proc Natl Acad Sci U S A. 1982 Dec;79(23):7595-7. doi: 10.1073/pnas.79.23.7595.
The L-alanine taste receptors of the channel catfish Ictalurus punctatus provide a useful biochemical model for studying taste receptor mechanisms. Mouse hybridomas that synthesize monoclonal antibodies have been produced. The antigen used to activate mouse spleen cells was the plasma membrane fraction obtained from the taste receptor-containing epithelium of the channel catfish. The spleen cells were fused with myeloma cells, Sp2/0-Ag14, to form hybridomas. To demonstrate inhibition of ligand binding by the product of these hybridomas, a catfish membrane fraction (fraction P2) was incubated with the antibody-containing preparation prior to assaying for L-[3H]alanine binding activity. We thereby demonstrated inhibition of binding of the taste ligand L-alanine to fraction P2. This approach should prove useful in further studies of receptor binding and transduction events in taste receptors.
斑点叉尾鮰的L-丙氨酸味觉受体为研究味觉受体机制提供了一个有用的生化模型。已经制备了合成单克隆抗体的小鼠杂交瘤。用于激活小鼠脾细胞的抗原是从斑点叉尾鮰含味觉受体的上皮细胞中获得的质膜部分。脾细胞与骨髓瘤细胞Sp2/0-Ag14融合形成杂交瘤。为了证明这些杂交瘤产物对配体结合的抑制作用，在测定L-[3H]丙氨酸结合活性之前，将鲶鱼膜部分（P2部分）与含抗体的制剂一起孵育。由此我们证明了味觉配体L-丙氨酸与P2部分的结合受到抑制。这种方法在味觉受体中受体结合和转导事件的进一步研究中应该是有用的。
