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Study on peptide-peptide interaction using high-performance affinity chromatography and quartz crystal microbalance biosensor

ISSN号：0251-0790
期刊名称：《高等学校化学学报》
时间：0
分类：Q516[生物学—生物化学]
作者机构：[1]Beijing National Laboratory for Molecular Sciences, Laboratory of Analytical Chemistry for Life Science, Institute of Chemistry,Chinese Academy of Sciences, Beijing 100080, China
相关基金：Supported by the National Natural Science Foundation of China （Grant Nos. 20575072, 20435030 and 90408018） and the Chinese Academy of Sciences （Grant No. KJCX2-SW-H06）The authors thank Dr. Zhong Yun of Yale University for critical reading of the manuscript.

作者： LUO Jia HUANG YanYan XIONG ShaoXiang LIU GuoQuan ZHAO Ruit[1]

关键词：肽-肽交互作用, 高效亲合色谱法, 石英晶体微平衡生物传感器, 人干扰素-β, antisense peptide, degeneracy, high-performance affinity chromatography, quartz crystal microbalance biosensor, human interferon-β

中文摘要：

在感觉和反感觉肽之间的特定的相互作用是学习 byhigh 表演亲密关系层析(HPAC ) 和石英水晶微量天秤(QCM ) 生物传感器。当感觉肽和它的 threeantisense 肽(AS-IFN 1， AS-IFN 2，和 AS-IFN 3 ) 根据遗传密码的退化被设计，人的干扰素贝它(hlFN 贝它) 的碎片 1-14 被选择。亲密关系列作为配位体和亲密关系与感觉肽被准备色析法的行为被评估。代替 Glu 的反感觉肽(AS-IFN 3 ) 在酸碱值显示出最强壮的绑定到使不能调动的感觉肽 7.5。一个石英水晶 microbalance-flowinjection 分析(QCM-FIA ) 系统被介绍调查识别进程 inreal 时间。在感觉肽和 AS-IFN 1 之间的平衡解离常数， AS-IFN 2 andAS-IFN 3 测量 2.08x10 ~(-4), 1.31x10 ~(-4) 和 2.22x10 ~(-5) mol/L 分别地。机制学习显示在感觉肽和 AS-IFN 3 之间的特定的识别是到期的 tosequence 依赖、多模式的亲密关系相互作用。

英文摘要：

The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography （HPAC） and quartz crystal microbalance （QCM） biosensor. Fragment 1-14 of human interferon-β （hlFN-βwas chosen as sense peptide and its three antisense peptides （AS-IFN 1, AS-IFN 2, and AS-IFN 3） were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide （AS-IFN 3） showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis （QCM-FIA） system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10^-4, 1.31×10^-4 and 2.22×10^-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.