Optimization of xenon biosensors for detection of protein interactions
Optimization of xenon biosensors for detection of protein interactions
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DOI：
10.1002/cbic.200500327
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发表时间：
2006-01-01
期刊：
影响因子：
3.2
通讯作者：
Wemmer, DE
中科院分区：
文献类型：
--
作者：
Lowery, TJ;Garcia, S;Wemmer, DE
超极化129 NMR光谱可以通过氙生物传感器检测特定低浓度生物分子分析物的存在，该氙生物传感器由封装氙的水溶性靶向cryptophane-A笼组成。在这项工作中，我们使用的原型生物素化氙生物传感器，以确定氙生物传感器的分子组成和蛋白质结合的共振特性之间的关系。非对映体重叠，偶极偶极耦合，化学位移各向异性，氙交换，和生物传感器构象交换上的蛋白质结合的生物传感器信号的影响进行了评估。结果发现，最佳的蛋白质结合的生物传感器信号可以通过最小化的生物传感器非对映异构体的数量，并使用适当长度的柔性接头。氙生物传感器的线宽和对蛋白质结合的化学位移的灵敏度两者被发现与连接体长度成反比。
Hyperpolarized Xe-129 NMR spectroscopy can detect the presence of specific low-concentration biomolecular analytes by means of a xenon biosensor that consists of a water-soluble, targeted cryptophane-A cage that encapsulates the xenon. In this work, we use the prototypical biotinylated xenon biosensor to determine the relationship between the molecular composition of the xenon biosensor and the characteristics of protein-bound resonances. The effects of diastereomer overlap, dipole-dipole coupling, chemical-shift anisotropy, xenon exchange, and biosensor conformotional exchange on the protein-bound biosensor signal were assessed. It was found that an optimal protein-bound biosensor signal can be obtained by minimizing the number of biosensor diastereomers and using a flexible linker of appropriate length. Both the line width and sensitivity of chemical shift to protein binding of the xenon biosensor were found to be inversely proportional to linker length.