A genome-inspired DNA ligand for the affinity capture of insulin and insulin-like growth factor-2.
A genome-inspired DNA ligand for the affinity capture of insulin and insulin-like growth factor-2.
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DOI：
10.1002/jssc.200900060
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发表时间：
2009-05
影响因子：
3.1
通讯作者：
McGown, Linda B.
中科院分区：
文献类型：
--
作者：
Xiao, Junfeng;Carter, Jennifer A.;Frederick, Kimberley A.;McGown, Linda B.
关键词：
人胰岛素基因的胰岛素连锁多态区（ILPR）包含相似的富g序列的串联重复序列，其中一些在体外形成分子内g四重结构。先前的研究表明，胰岛素与ILPR变体a形成的分子内g -四重体具有亲和力结合。本文报告了胰岛素和高度同源的胰岛素样生长因子2 （IGF-2）与ILPR变体a、h和i的相互作用。圆二色性表明，变体a和h形成了分子内g -四重体。亲和MALDI质谱和表面等离子体共振用于比较蛋白质捕获和结合强度。胰岛素和IGF-2对变异a和h表现出高结合亲和力，而对i则没有，这表明分子内g -四联体的参与。胰岛素与变异a之间的相互作用是独特的，它与KD~10−13 M和KD~10−7 M存在两种结合相互作用，而与变异h （KD~10−8 M）的胰岛素或与任何一种变异（KD的~10−9 D M）的IGF-2没有观察到这种相互作用。该结果为胰岛素和IGF-2的DNA结合配体的设计提供了基础，并为基于基因组启发序列发现DNA亲和结合配体提供了新途径，而不是传统的组合选择途径来发现适体。
The insulin-linked polymorphic region (ILPR) of the human insulin gene contains tandem repeats of similar G-rich sequences, some of which form intramolecular G-quadruplex structures in vitro. Previous work showed affinity binding of insulin to an intramolecular G-quadruplex formed by ILPR variant a. Here we report on interactions of insulin and the highly homologous insulin-like growth factor 2 (IGF-2) with ILPR variants a, h and i. Circular dichroism indicated intramolecular G-quadruplex formation for variants a and h. Affinity MALDI mass spectrometry and surface plasmon resonance were used to compare protein capture and binding strengths. Insulin and IGF-2 exhibited high binding affinity for variants a and h but not i, indicating the involvement of intramolecular G-quadruplexes. Interaction between insulin and variant a was unique in the appearance of two binding interactions with KD~10−13 M and KD~10−7 M, which was not observed for insulin with variant h (KD~10−8 M) or IGF-2 with either variant (KD’s~10−9 D M). The results provide a basis for design of DNA binding ligands for insulin and IGF-2 and support a new approach to discovery of DNA affinity binding ligands based on genome-inspired sequences rather than the traditional combinatorial selection route to aptamer discovery.
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影响因子：
2
作者：
Gray, Donald M.;Wen, Jin-Der;Fleischhauer, Joerg
通讯作者：
Fleischhauer, Joerg
影响因子：
7.4
作者：
Cole, Jacquelyn R.;Dick, Lawrence W., Jr.;McGown, Linda B.
通讯作者：
McGown, Linda B.
影响因子：
3.5
作者：
HAMMONDKOSACK, MCU;DOCHERTY, K
通讯作者：
DOCHERTY, K
影响因子：
7.7
作者：
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通讯作者：
Gabbay, KH
影响因子：
14.9
作者：
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通讯作者：
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