Engineering of insulin receptor isoform-selective insulin analogues.
Engineering of insulin receptor isoform-selective insulin analogues.
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DOI：
10.1371/journal.pone.0020288
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Kjeldsen T
中科院分区：
文献类型：
--
作者：
Glendorf T;Stidsen CE;Norrman M;Nishimura E;Sørensen AR;Kjeldsen T
胰岛素受体(IR)以A和B两种亚型存在，并且这种亚型的表达模式具有组织特异性。胰岛素B链的C-末端对于受体结合是重要的，并且已被证明与胰岛素受体接触，恰好与A和B异构体不同的区域相邻。本研究的目的是研究B链的C末端在IR异构体结合中的重要性，以探索设计组织特异性/肝脏特异性胰岛素类似物的可能性。通过总氨基酸扫描诱变构建胰岛素类似物文库。用闪烁邻近分析技术进行竞争分析，确定IR的A和B亚型的相对结合亲和力。用X-射线单晶衍射法获得了晶体的结构信息。B25A或B25N突变的引入导致类似物对B的偏好是A异构体的2倍，而B25Y替换则相反。B27位的酸性氨基酸残基使带有B25N突变的类似物对B受体亚型的亲和力增加了2倍。此外，B25H与B27D或B27E的结合也导致B异构体偏好类似物(2倍偏好)，即使相应的单一突变类似物在相对异构体结合亲和力上没有差异。我们发现了一类新的IR异构体选择性胰岛素类似物，与人胰岛素相比，IR的A或B异构体的相对结合亲和力相差2-4倍。我们的结果表明，胰岛素分子中B25位的突变或与B27位的突变一起提供IR异构体的选择性。异构体优先类似物可能为开发具有更好临床益处的胰岛素类似物提供新的机会。
The insulin receptor (IR) exists in two isoforms, A and B, and the isoform expression pattern is tissue-specific. The C-terminus of the insulin B chain is important for receptor binding and has been shown to contact the IR just adjacent to the region where the A and B isoforms differ. The aim of this study was to investigate the importance of the C-terminus of the B chain in IR isoform binding in order to explore the possibility of engineering tissue-specific/liver-specific insulin analogues. Insulin analogue libraries were constructed by total amino acid scanning mutagenesis. The relative binding affinities for the A and B isoform of the IR were determined by competition assays using scintillation proximity assay technology. Structural information was obtained by X-ray crystallography. Introduction of B25A or B25N mutations resulted in analogues with a 2-fold preference for the B compared to the A isoform, whereas the opposite was observed with a B25Y substitution. An acidic amino acid residue at position B27 caused an additional 2-fold selective increase in affinity for the receptor B isoform for analogues bearing a B25N mutation. Furthermore, the combination of B25H with either B27D or B27E also resulted in B isoform-preferential analogues (2-fold preference) even though the corresponding single mutation analogues displayed no differences in relative isoform binding affinity. We have discovered a new class of IR isoform-selective insulin analogues with 2–4-fold differences in relative binding affinities for either the A or the B isoform of the IR compared to human insulin. Our results demonstrate that a mutation at position B25 alone or in combination with a mutation at position B27 in the insulin molecule confers IR isoform selectivity. Isoform-preferential analogues may provide new opportunities for developing insulin analogues with improved clinical benefits.
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影响因子：
64.8
作者：
HUA, QX;SHOELSON, SE;WEISS, MA
通讯作者：
WEISS, MA
DOI：
10.1515/bchm3.1985.366.1.521
发表时间：
1985-01-01
期刊：
BIOLOGICAL CHEMISTRY HOPPE-SEYLER
影响因子：
--
作者：
FISCHER, WH;SAUNDERS, D;ZAHN, H
通讯作者：
ZAHN, H
DOI：
10.1098/rstb.1988.0058
发表时间：
1988-07-06
影响因子：
6.3
作者：
BAKER, EN;BLUNDELL, TL;VIJAYAN, NM
通讯作者：
VIJAYAN, NM
影响因子：
2.9
作者：
KAARSHOLM, NC;NORRIS, K;HAVELUND, S
通讯作者：
HAVELUND, S
影响因子：
2.9
作者：
NAKAGAWA, SH;TAGER, HS
通讯作者：
TAGER, HS