High affinity insulin binding by soluble insulin receptor extracellular domain fused to a leucine zipper
High affinity insulin binding by soluble insulin receptor extracellular domain fused to a leucine zipper
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DOI：
10.1016/s0014-5793(00)01872-x
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发表时间：
2000-08-11
期刊：
影响因子：
3.5
通讯作者：
Ward, CW
中科院分区：
文献类型：
--
作者：
Hoyne, PA;Cosgrove, LJ;Ward, CW
在胞外域末端被截短的胰岛素受体（IR）形成二聚体，其后胰岛素具有与野生型受体不同的特征。与全长IR相比，这些可溶性IR对胰岛素的亲和力降低，并且与用全长IR、在跨膜区的C末端截短的IR和融合到胞外域的IR胞外域获得的曲线图相比，表现出线性斯卡查德图。来自 Pc 或 lambda 免疫球蛋白的自缔合恒定域。在本报告中，我们将 1R 胞外域与来自酿酒酵母转录激活剂 GCN4 的 33 残基亮氨酸拉链融合。这种融合蛋白以与天然受体相当的方式以高亲和力结合胰岛素。 hlRedZip 各自的解离常数为 K-d1 8.2 x 10(-11) M 和 K-d2 1.6 x 10(-8) M，膜锚定天然受体的解离常数为 K-d1 5.7 x 10(-11) 和 K-d2 6.3 x 10(-9) M。 (C) 2000 年欧洲生化学会联合会。由 Elsevier Science B.V. 出版。保留所有权利。
Insulin receptors (IRs) that are truncated at the end of the ectodomain form dimers that hind insulin with different characteristics to wild type receptors, These soluble IRs have lowered affinity for insulin compared with full-length IR, and exhibit linear Scatchard plots in contrast to the curvilinear plots obtained with full-length IR, IR truncated at the C-terminus of the transmembrane region and IR ectodomains fused to the self-associating constant domains from Pc or lambda immunoglobulins. In this report, we have fused the 1R ectodomain to the 33 residue leucine zipper from the transcriptional activator GCN4 of Saccharomyces cerevisiae. This fusion protein binds insulin with high affinity in a manner comparable to native receptor. The respective dissociation constants were K-d1 8.2 x 10(-11) M and K-d2 1.6 x 10(-8) M for hlRedZip and K-d1 5.7 x 10(-11) and K-d2 6.3 x 10(-9) M for membrane-anchored, native receptor. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.