Buggy J, Rossomando A, MacDougall M, Mierz D, Wunderlich D, Yoo-Warren H
Institute for Research Technologies, Mayer, Inc., West Haven, Connecticut, USA.
Horm Metab Res. 1996 May;28(5):215-9. doi: 10.1055/s-2007-979167.
This paper describes the development and characterization of the first monoclonal antibody specific for the recently cloned human glucagon receptor (hGR), and its use in probing receptor structure and function. We demonstrate specificity of one of the antibodies, CIV395.7A, by immunofluorescence staining and immunoprecipitation analysis. In addition, CIV395.7A specifically competes with glucagon for the hormone binding site on the receptor, indicating that the antibody's specific recognition epitope overlaps with the receptor's hormone binding domain. As a consequence, the mAB antagonizes glucagon-stimulated signal transduction as assayed by in vitro cAMP accumulation. Binding inhibition studies further reveal that the antibody specifically recognizes the human and rat GR, but not mouse. Using hGR/glucagon-like peptide I receptor chimeras, we have localized the recognition epitope of the antibody to the membrane-proximal half of the amino-terminal extension of the receptor, thus defining a domain on the receptor which is involved in glucagon binding.
本文描述了针对最近克隆的人胰高血糖素受体（hGR）的首个单克隆抗体的研制与特性鉴定，以及该抗体在探究受体结构与功能方面的应用。我们通过免疫荧光染色和免疫沉淀分析，证实了其中一种抗体CIV395.7A的特异性。此外，CIV395.7A能与胰高血糖素在受体上的激素结合位点特异性竞争，这表明该抗体的特异性识别表位与受体的激素结合域重叠。因此，如通过体外环磷酸腺苷（cAMP）积累检测所示，该单克隆抗体可拮抗胰高血糖素刺激的信号转导。结合抑制研究进一步表明，该抗体能特异性识别人和大鼠的GR，但不能识别小鼠的GR。利用hGR/胰高血糖素样肽I受体嵌合体，我们已将该抗体的识别表位定位到受体氨基末端延伸区靠近膜的一半，从而确定了受体上一个参与胰高血糖素结合的结构域。
