Rojas F J, Swartz T L, Iyengar R, Garber A J, Birnbaumer L
Endocrinology. 1983 Aug;113(2):711-9. doi: 10.1210/endo-113-2-711.
The synthesis of [125I-Tyr10]monoiodoglucagon from glucagon and carrier-free 125I using 1,3,4,6-tetrachloro-3-6-diphenylglycouril (Iodogen) and its separation in pure form by reverse phase high pressure liquid chromatography (HPLC) over C18-muBondapak columns using two consecutive linear gradients between solvent A [40:60 mixture of methanol and 10 mM H3PO4 in H2O (pH adjusted to 3.0 with triethylamine)] and solvent B (50:50 mixture of acetonitrile and 0.1 M Tris-HCl, pH 9.0) is reported. The newly synthesized [125I]monoiodoglucagon is shown to activate adenylyl cyclase in liver membranes with an EC50 between 5- and 8-fold lower than that of native glucagon. Further, it binds specifically to sites on liver plasma membranes that have the characteristics of glucagon receptors in terms of guanine nucleotide sensitivity and rates of reaction. It is suggested that [125I-Tyr10]monoiodoglucagon is a suitable probe for studying structural and functional properties of glucagon receptors.
报道了使用1,3,4,6-四氯-3,6-二苯基甘脲（碘甘脲）由胰高血糖素和无载体125I合成[125I-Tyr10]单碘胰高血糖素，并通过反相高压液相色谱（HPLC）在C18-μBondapak柱上使用溶剂A[甲醇与10 mM H3PO4在水中的40:60混合物（用三乙胺将pH调节至3.0）]和溶剂B（乙腈与0.1 M Tris-HCl的50:50混合物，pH 9.0）之间的两个连续线性梯度将其分离成纯形式。新合成的[125I]单碘胰高血糖素显示出在肝细胞膜中激活腺苷酸环化酶，其半数有效浓度（EC50）比天然胰高血糖素低5至8倍。此外，它特异性结合肝细胞膜上具有胰高血糖素受体在鸟嘌呤核苷酸敏感性和反应速率方面特征的位点。有人认为[125I-Tyr10]单碘胰高血糖素是研究胰高血糖素受体结构和功能特性的合适探针。
